Cyclopropyl modulators of p2y12 receptor

ABSTRACT

The present invention relates to new cyclopropyl modulators of P2Y12 receptor activity, pharmaceutical compositions thereof, and methods of use thereof.

This application is a division of U.S. patent application Ser. No.12/844,017, filed Jul. 27, 2010, which claims the benefit of priority ofU.S. provisional application No. 61/228,913, filed Jul. 27, 2009, thedisclosures of which are hereby incorporated by reference as if writtenherein in their entireties.

Disclosed herein are new substituted cyclopropyl compounds,pharmaceutical compositions made thereof, and methods to modulate P2Y12receptor activity in a subject are also provided for, for the treatmentof disorders such as arterial thrombosis and coronary artery disease.

Ticagrelor (AR-C126532, AZD-6140, Brilinta®, CAS #274693-27-5),3-[7-[[(1R,2S)-2-(3,4-difluorophenyl)cyclopropyl]amino]-5-(propylthio)-3H-1,2,3-triazolo[4,5-d]pyrimidin-3-yl]-5-(2-hydroxyethoxy)-(1S,2S,3R,5S)-1,2-cyclopentanediol,is a P2Y12 receptor antagonist. Ticagrelor is currently underinvestigation for the treatment of arterial thrombosis (Tantry et al.,Exp. Opin. Invest. Drugs 2007, 16(2), 225-229; Husted et al., Eur. HeartJ. 2006, 27(9), 1038-1047; and WO 2000034283). Ticagrelor has also shownpromise in treating coronary artery disease and other disorders relatedto platelet aggregation (Tantry et al., Exp. Opin. Invest. Drugs 2007,16(2), 225-229; Husted et al., Eur. Heart J. 2006, 27(9), 1038-1047; andWO 2000034283).

Ticagrelor is subject to CYP450-mediated oxidative metabolism, formingan active metabolite AR-C124910XX (Husted et al., Eur. Heart J. 2006,27, 1038-1047). Adverse effects associated with ticagrelor includeexcessive bleeding.

Deuterium Kinetic Isotope Effect

In order to eliminate foreign substances such as therapeutic agents, theanimal body expresses various enzymes, such as the cytochrome P₄₅₀enzymes (CYPs), esterases, proteases, reductases, dehydrogenases, andmonoamine oxidases, to react with and convert these foreign substancesto more polar intermediates or metabolites for renal excretion. Suchmetabolic reactions frequently involve the oxidation of acarbon-hydrogen (C—H) bond to either a carbon-oxygen (C—O) or acarbon-carbon (C—C) 2π-bond. The resultant metabolites may be stable orunstable under physiological conditions, and can have substantiallydifferent pharmacokinetic, pharmacodynamic, and acute and long-termtoxicity profiles relative to the parent compounds. For most drugs, suchoxidations are generally rapid and ultimately lead to administration ofmultiple or high daily doses.

The relationship between the activation energy and the rate of reactionmay be quantified by the Arrhenius equation, k=Ae^(−Eact/RT). TheArrhenius equation states that, at a given temperature, the rate of achemical reaction depends exponentially on the activation energy(E_(act)).

The transition state in a reaction is a short lived state along thereaction pathway during which the original bonds have stretched to theirlimit. By definition, the activation energy E_(act) for a reaction isthe energy required to reach the transition state of that reaction. Oncethe transition state is reached, the molecules can either revert to theoriginal reactants, or form new bonds giving rise to reaction products.A catalyst facilitates a reaction process by lowering the activationenergy leading to a transition state. Enzymes are examples of biologicalcatalysts.

Carbon-hydrogen bond strength is directly proportional to the absolutevalue of the ground-state vibrational energy of the bond. Thisvibrational energy depends on the mass of the atoms that form the bond,and increases as the mass of one or both of the atoms making the bondincreases. Since deuterium (D) has twice the mass of protium (¹H), a C-Dbond is stronger than the corresponding C—¹H bond. If a C—¹H bond isbroken during a rate-determining step in a chemical reaction (i.e. thestep with the highest transition state energy), then substituting adeuterium for that protium will cause a decrease in the reaction rate.This phenomenon is known as the Deuterium Kinetic Isotope Effect (DKIE).The magnitude of the DKIE can be expressed as the ratio between therates of a given reaction in which a C—¹H bond is broken, and the samereaction where deuterium is substituted for protium. The DKIE can rangefrom about 1 (no isotope effect) to very large numbers, such as 50 ormore. Substitution of tritium for hydrogen results in yet a strongerbond than deuterium and gives numerically larger isotope effectsDeuterium (²H or D) is a stable and non-radioactive isotope of hydrogenwhich has approximately twice the mass of protium (¹H), the most commonisotope of hydrogen. Deuterium oxide (D₂O or “heavy water”) looks andtastes like H₂O, but has different physical properties.

When pure D₂O is given to rodents, it is readily absorbed. The quantityof deuterium required to induce toxicity is extremely high. When about0-15% of the body water has been replaced by D₂O, animals are healthybut are unable to gain weight as fast as the control (untreated) group.When about 15-20% of the body water has been replaced with D₂O, theanimals become excitable. When about 20-25% of the body water has beenreplaced with D₂O, the animals become so excitable that they go intofrequent convulsions when stimulated. Skin lesions, ulcers on the pawsand muzzles, and necrosis of the tails appear. The animals also becomevery aggressive. When about 30% of the body water has been replaced withD₂O, the animals refuse to eat and become comatose. Their body weightdrops sharply and their metabolic rates drop far below normal, withdeath occurring at about 30 to about 35% replacement with D₂O. Theeffects are reversible unless more than thirty percent of the previousbody weight has been lost due to D₂O, Studies have also shown that theuse of D₂O can delay the growth of cancer cells and enhance thecytotoxicity of certain antineoplastic agents.

Deuteration of pharmaceuticals to improve pharmacokinetics (PK),pharmacodynamics (PD), and toxicity profiles has been demonstratedpreviously with some classes of drugs. For example, the DKIE was used todecrease the hepatotoxicity of halothane, presumably by limiting theproduction of reactive species such as trifluoroacetyl chloride.However, this method may not be applicable to all drug classes. Forexample, deuterium incorporation can lead to metabolic switching.Metabolic switching occurs when xenogens, sequestered by Phase Ienzymes, bind transiently and re-bind in a variety of conformationsprior to the chemical reaction (e.g., oxidation). Metabolic switching isenabled by the relatively vast size of binding pockets in many Phase Ienzymes and the promiscuous nature of many metabolic reactions.Metabolic switching can lead to different proportions of knownmetabolites as well as altogether new metabolites. This new metabolicprofile may impart more or less toxicity. Such pitfalls are non-obviousand are not predictable a priori for any drug class.

Ticagrelor is a P2Y12 receptor antagonist. The carbon-hydrogen bonds ofticagrelor contain a naturally occurring distribution of hydrogenisotopes, namely ¹H or protium (about 99.9844%), ²H or deuterium (about0.0156%), and ³H or tritium (in the range between about 0.5 and 67tritium atoms per 10¹⁸ protium atoms). Increased levels of deuteriumincorporation may produce a detectable Deuterium Kinetic Isotope Effect(DKIE) that could effect the pharmacokinetic, pharmacologic and/ortoxicologic profiles of ticagrelor in comparison with ticagrelor havingnaturally occurring levels of deuterium.

Based on discoveries made in our laboratory, as well as considering theliterature, ticagrelor is likely metabolized in humans at the2-hydroxyethoxy group, the S-propyl group, and the cyclopropyl group.The current approach has the potential to prevent metabolism at thesesites. Other sites on the molecule may also undergo transformationsleading to metabolites with as-yet-unknown pharmacology/toxicology.Limiting the production of these metabolites has the potential todecrease the danger of the administration of such drugs and may evenallow increased dosage and/or increased efficacy. All of thesetransformations can occur through polymorphically-expressed enzymes,exacerbating interpatient variability. Further, some disorders are besttreated when the subject is medicated around the clock or for anextended period of time. For all of the foregoing reasons, a medicinewith a longer half-life may result in greater efficacy and cost savings.Various deuteration patterns can be used to (a) reduce or eliminateunwanted metabolites, (b) increase the half-life of the parent drug, (c)decrease the number of doses needed to achieve a desired effect, (d)decrease the amount of a dose needed to achieve a desired effect, (e)increase the formation of active metabolites, if any are formed, (f)decrease the production of deleterious metabolites in specific tissues,and/or (g) create a more effective drug and/or a safer drug forpolypharmacy, whether the polypharmacy be intentional or not. Thedeuteration approach has the strong potential to slow the metabolism ofticagrelor and attenuate interpatient variability.

Novel compounds and pharmaceutical compositions, certain of which havebeen found to modulate P2Y12 receptor activity have been discovered,together with methods of synthesizing and using the compounds, includingmethods for the treatment of P2Y12 receptor-mediated disorders in apatient by administering the compounds as disclosed herein.

In certain embodiments of the present invention, compounds havestructural Formula I:

or a pharmaceutically acceptable salt thereof, wherein:

R₁-R₂₈ are independently selected from the group consisting of hydrogenand deuterium; and

at least one of R₁-R₂₈ is deuterium.

Certain compounds disclosed herein may possess useful P2Y12 receptormodulating activity, and may be used in the treatment or prophylaxis ofa disorder in which P2Y12 receptors play an active role. Thus, certainembodiments also provide pharmaceutical compositions comprising one ormore compounds disclosed herein together with a pharmaceuticallyacceptable carrier, as well as methods of making and using the compoundsand compositions. Certain embodiments provide methods for modulatingP2Y12 receptor activity. Other embodiments provide methods for treatinga P2Y12 receptor-mediated disorder in a patient in need of suchtreatment, comprising administering to said patient a therapeuticallyeffective amount of a compound or composition according to the presentinvention. Also provided is the use of certain compounds disclosedherein for use in the manufacture of a medicament for the prevention ortreatment of a disorder ameliorated by modulating P2Y12 receptoractivity.

The compounds as disclosed herein may also contain less prevalentisotopes for other elements, including, but not limited to, ¹³C or ¹⁴Cfor carbon, ³³S, ³⁴S, or ³⁶S for sulfur, ¹⁵N for nitrogen, and ¹⁷O or¹⁸O for oxygen.

In certain embodiments, the compound disclosed herein may expose apatient to a maximum of about 0.000005% D₂O or about 0.00001% DHO,assuming that all of the C-D bonds in the compound as disclosed hereinare metabolized and released as D₂O or DHO. In certain embodiments, thelevels of D₂O shown to cause toxicity in animals is much greater thaneven the maximum limit of exposure caused by administration of thedeuterium enriched compound as disclosed herein. Thus, in certainembodiments, the deuterium-enriched compound disclosed herein should notcause any additional toxicity due to the formation of D₂O or DHO upondrug metabolism.

In certain embodiments, the deuterated compounds disclosed hereinmaintain the beneficial aspects of the corresponding non-isotopicallyenriched molecules while substantially increasing the maximum tolerateddose, decreasing toxicity, increasing the half-life (T_(1/2)), loweringthe maximum plasma concentration (C_(max)) of the minimum efficaciousdose (MED), lowering the efficacious dose and thus decreasing thenon-mechanism-related toxicity, and/or lowering the probability ofdrug-drug interactions.

All publications and references cited herein are expressly incorporatedherein by reference in their entirety. However, with respect to anysimilar or identical terms found in both the incorporated publicationsor references and those explicitly put forth or defined in thisdocument, then those terms definitions or meanings explicitly put forthin this document shall control in all respects.

As used herein, the terms below have the meanings indicated.

The singular forms “a”, “an”, and “the” may refer to plural articlesunless specifically stated otherwise.

The term “about”, as used herein, is intended to qualify the numericalvalues which it modifies, denoting such a value as variable within amargin of error. When no particular margin of error, such as a standarddeviation to a mean value given in a chart or table of data, is recited,the term “about” should be understood to mean that range which wouldencompass the recited value and the range which would be included byrounding up or down to that figure as well, taking into accountsignificant figures.

When ranges of values are disclosed, and the notation “from n₁ . . . ton₂” or “n₁-n₂” is used, where n₁ and n₂ are the numbers, then unlessotherwise specified, this notation is intended to include the numbersthemselves and the range between them. This range may be integral orcontinuous between and including the end values.

The term “deuterium enrichment” refers to the percentage ofincorporation of deuterium at a given position in a molecule in theplace of hydrogen. For example, deuterium enrichment of 1% at a givenposition means that 1% of molecules in a given sample contain deuteriumat the specified position. Because the naturally occurring distributionof deuterium is about 0.0156%, deuterium enrichment at any position in acompound synthesized using non-enriched starting materials is about0.0156%. The deuterium enrichment can be determined using conventionalanalytical methods known to one of ordinary skill in the art, includingmass spectrometry and nuclear magnetic resonance spectroscopy.

The term “is/are deuterium”, when used to describe a given position in amolecule such as R₁-R₂₈ or the symbol “D”, when used to represent agiven position in a drawing of a molecular structure, means that thespecified position is enriched with deuterium above the naturallyoccurring distribution of deuterium. In one embodiment deuteriumenrichment is no less than about 1%, in another no less than about 5%,in another no less than about 10%, in another no less than about 20%, inanother no less than about 50%, in another no less than about 70%, inanother no less than about 80%, in another no less than about 90%, or inanother no less than about 98% of deuterium at the specified position.

The term “isotopic enrichment” refers to the percentage of incorporationof a less prevalent isotope of an element at a given position in amolecule in the place of the more prevalent isotope of the element.

The term “non-isotopically enriched” refers to a molecule in which thepercentages of the various isotopes are substantially the same as thenaturally occurring percentages.

Asymmetric centers exist in the compounds disclosed herein. Thesecenters are designated by the symbols “R” or “S”, depending on theconfiguration of substituents around the chiral carbon atom. It shouldbe understood that the invention encompasses all stereochemical isomericforms, including diastereomeric, enantiomeric, and epimeric forms, aswell as D-isomers and L-isomers, and mixtures thereof. Individualstereoisomers of compounds can be prepared synthetically fromcommercially available starting materials which contain chiral centersor by preparation of mixtures of enantiomeric products followed byseparation such as conversion to a mixture of diastereomers followed byseparation or recrystallization, chromatographic techniques, directseparation of enantiomers on chiral chromatographic columns, or anyother appropriate method known in the art. Starting compounds ofparticular stereochemistry are either commercially available or can bemade and resolved by techniques known in the art. Additionally, thecompounds disclosed herein may exist as geometric isomers. The presentinvention includes all cis, trans, syn, anti, entgegen (E), and zusammen(Z) isomers as well as the appropriate mixtures thereof. Additionally,compounds may exist as tautomers; all tautomeric isomers are provided bythis invention. Additionally, the compounds disclosed herein can existin unsolvated as well as solvated forms with pharmaceutically acceptablesolvents such as water, ethanol, and the like. In general, the solvatedforms are considered equivalent to the unsolvated forms.

The term “bond” refers to a covalent linkage between two atoms, or twomoieties when the atoms joined by the bond are considered to be part oflarger substructure. A bond may be single, double, or triple unlessotherwise specified. A dashed line between two atoms in a drawing of amolecule indicates that an additional bond may be present or absent atthat position.

The term “disorder” as used herein is intended to be generallysynonymous, and is used interchangeably with, the terms “disease”,“syndrome”, and “condition” (as in medical condition), in that allreflect an abnormal condition of the human or animal body or of one ofits parts that impairs normal functioning, is typically manifested bydistinguishing signs and symptoms.

The terms “treat”, “treating”, and “treatment” are meant to includealleviating or abrogating a disorder or one or more of the symptomsassociated with a disorder; or alleviating or eradicating the cause(s)of the disorder itself. As used herein, reference to “treatment” of adisorder is intended to include prevention. The terms “prevent”,“preventing”, and “prevention” refer to a method of delaying orprecluding the onset of a disorder; and/or its attendant symptoms,barring a subject from acquiring a disorder or reducing a subject's riskof acquiring a disorder.

The term “therapeutically effective amount” refers to the amount of acompound that, when administered, is sufficient to prevent developmentof, or alleviate to some extent, one or more of the symptoms of thedisorder being treated. The term “therapeutically effective amount” alsorefers to the amount of a compound that is sufficient to elicit thebiological or medical response of a cell, tissue, system, animal, orhuman that is being sought by a researcher, veterinarian, medicaldoctor, or clinician.

The term “subject” refers to an animal, including, but not limited to, aprimate (e.g., human, monkey, chimpanzee, gorilla, and the like),rodents (e.g., rats, mice, gerbils, hamsters, ferrets, and the like),lagomorphs, swine (e.g., pig, miniature pig), equine, canine, feline,and the like. The terms “subject” and “patient” are used interchangeablyherein in reference, for example, to a mammalian subject, such as ahuman patient.

The term “combination therapy” means the administration of two or moretherapeutic agents to treat a therapeutic disorder described in thepresent disclosure. Such administration encompasses co-administration ofthese therapeutic agents in a substantially simultaneous manner, such asin a single capsule having a fixed ratio of active ingredients or inmultiple, separate capsules for each active ingredient. In addition,such administration also encompasses use of each type of therapeuticagent in a sequential manner. In either case, the treatment regimen willprovide beneficial effects of the drug combination in treating thedisorders described herein.

The term “P2Y12 receptor” refers to a G-protein coupled receptor locatedon the platelet membrane. The P2Y12 receptor (also known as P2T, P2YADP,or P2TAC) is primarily involved in mediating plateletaggregation/activation. The pharmacological characteristics of thisreceptor have been described, for example, by Humphries et al., Br. J.Pharmacology 1994, 113, 1057-1063; and Fagura et al., Br. J.Pharmacology 1998, 124, 157-164.

The term “P2Y12 receptor-mediated disorder”, refers to a disorder thatis characterized by abnormal P2Y12 receptor activity or excessiveplatelet aggregation, or normal platelet aggregation or normal P2Y12receptor activity that when modulated ameliorates other abnormalbiochemical processes. A P2Y12 receptor-mediated disorder may becompletely or partially mediated by modulating P2Y12 receptor activity.In particular, a P2Y12 receptor-mediated disorder is one in whichmodulation of P2Y12 receptor activity results in some effect on theunderlying disorder e.g., administration of a P2Y12 receptor modulatorresults in some improvement in at least some of the patients beingtreated.

The term “P2Y12 receptor modulator”, refers to the ability of a compounddisclosed herein to alter the function of P2Y12 receptors. A P2Y12receptor modulator may activate the activity of a P2Y12 receptor, mayactivate or inhibit the activity of a P2Y12 receptor depending on theconcentration of the compound exposed to the P2Y12 receptor, or mayinhibit the activity of a P2Y12 receptor. Such activation or inhibitionmay be contingent on the occurrence of a specific event, such asactivation of a signal transduction pathway, and/or may be manifest onlyin particular cell types. The term “P2Y12 receptor modulator”, alsorefers to altering the function of a P2Y12 receptor by increasing ordecreasing the probability that a complex forms between a P2Y12 receptorand a natural binding partner. A P2Y12 receptor modulator may increasethe probability that such a complex forms between the P2Y12 receptor andthe natural binding partner, may increase or decrease the probabilitythat a complex forms between the P2Y12 receptor and the natural bindingpartner depending on the concentration of the compound exposed to theP2Y12 receptor, and or may decrease the probability that a complex formsbetween the P2Y12 receptor and the natural binding partner. In someembodiments, modulation of the P2Y12 receptor activity may be assessedusing the method described in Husted et al., Eur. Heart J. 2006, 27(9),1038-1047; WO 2000034283; WO 199905142.

The term “therapeutically acceptable” refers to those compounds (orsalts, prodrugs, tautomers, zwitterionic forms, etc.) which are suitablefor use in contact with the tissues of patients without excessivetoxicity, irritation, allergic response, immunogenecity, arecommensurate with a reasonable benefit/risk ratio, and are effective fortheir intended use.

The term “pharmaceutically acceptable carrier”, “pharmaceuticallyacceptable excipient”, “physiologically acceptable carrier”, or“physiologically acceptable excipient” refers to apharmaceutically-acceptable material, composition, or vehicle, such as aliquid or solid filler, diluent, excipient, solvent, or encapsulatingmaterial. Each component must be “pharmaceutically acceptable” in thesense of being compatible with the other ingredients of a pharmaceuticalformulation. It must also be suitable for use in contact with the tissueor organ of humans and animals without excessive toxicity, irritation,allergic response, immunogenecity, or other problems or complications,commensurate with a reasonable benefit/risk ratio. See, Remington: TheScience and Practice of Pharmacy, 21st Edition; Lippincott Williams &Wilkins: Philadelphia, Pa., 2005; Handbook of Pharmaceutical Excipients,5th Edition; Rowe et al., Eds., The Pharmaceutical Press and theAmerican Pharmaceutical Association: 2005; and Handbook ofPharmaceutical Additives, 3rd Edition; Ash and Ash Eds., GowerPublishing Company: 2007; Pharmaceutical Preformulation and Formulation,Gibson Ed., CRC Press LLC: Boca Raton, Fla., 2004).

The terms “active ingredient”, “active compound”, and “active substance”refer to a compound, which is administered, alone or in combination withone or more pharmaceutically acceptable excipients or carriers, to asubject for treating, preventing, or ameliorating one or more symptomsof a disorder.

The terms “drug”, “therapeutic agent”, and “chemotherapeutic agent”refer to a compound, or a pharmaceutical composition thereof, which isadministered to a subject for treating, preventing, or ameliorating oneor more symptoms of a disorder.

The term “release controlling excipient” refers to an excipient whoseprimary function is to modify the duration or place of release of theactive substance from a dosage form as compared with a conventionalimmediate release dosage form.

The term “nonrelease controlling excipient” refers to an excipient whoseprimary function do not include modifying the duration or place ofrelease of the active substance from a dosage form as compared with aconventional immediate release dosage form.

The term “prodrug” refers to a compound functional derivative of thecompound as disclosed herein and is readily convertible into the parentcompound in vivo. Prodrugs are often useful because, in some situations,they may be easier to administer than the parent compound. They may, forinstance, be bioavailable by oral administration whereas the parentcompound is not. The prodrug may also have enhanced solubility inpharmaceutical compositions over the parent compound. A prodrug may beconverted into the parent drug by various mechanisms, includingenzymatic processes and metabolic hydrolysis. See Harper, Progress inDrug Research 1962, 4, 221-294; Morozowich et al. in “Design ofBiopharmaceutical Properties through Prodrugs and Analogs,” Roche Ed.,APHA Acad. Pharm. Sci. 1977; “Bioreversible Carriers in Drug in DrugDesign, Theory and Application,” Roche Ed., APHA Acad. Pharm. Sci. 1987;“Design of Prodrugs,” Bundgaard, Elsevier, 1985; Wang et al., Curr.Pharm. Design 1999, 5, 265-287; Pauletti et al., Adv. Drug. DeliveryRev. 1997, 27, 235-256; Mizen et al., Pharm. Biotech. 1998, 11, 345-365;Gaignault et al., Pract. Med. Chem. 1996, 671-696; Asgharnejad in“Transport Processes in Pharmaceutical Systems,” Amidon et al., Ed.,Marcell Dekker, 185-218, 2000; Balant et al., Eur. J. Drug Metab.Pharmacokinet. 1990, 15, 143-53; Balimane and Sinko, Adv. Drug DeliveryRev. 1999, 39, 183-209; Browne, Clin. Neuropharmacol. 1997, 20, 1-12;Bundgaard, Arch. Pharm. Chem. 1979, 86, 1-39; Bundgaard, Controlled DrugDelivery 1987, 17, 179-96; Bundgaard, Adv. Drug Delivery Rev. 1992, 8,1-38; Fleisher et al., Adv. Drug Delivery Rev. 1996, 19, 115-130;Fleisher et al., Methods Enzymol. 1985, 112, 360-381; Farquhar et al.,J. Pharm. Sci. 1983, 72, 324-325; Freeman et al., J. Chem. Soc., Chem.Commun. 1991, 875-877; Friis and Bundgaard, Eur. J. Pharm. Sci. 1996, 4,49-59; Gangwar et al., Des. Biopharm. Prop. Prodrugs Analogs, 1977,409-421; Nathwani and Wood, Drugs 1993, 45, 866 94; Sinhababu andThakker, Adv. Drug Delivery Rev. 1996, 19, 241-273; Stella et al., Drugs1985, 29, 455-73; Tan et al., Adv. Drug Delivery Rev. 1999, 39, 117-151;Taylor, Adv. Drug Delivery Rev. 1996, 19, 131-148; Valentino andBorchardt, Drug Discovery Today 1997, 2, 148-155; Wiebe and Knaus, Adv.Drug Delivery Rev. 1999, 39, 63-80; Waller et al., Br. J. Clin. Pharmac.1989, 28, 497-507.

The compounds disclosed herein can exist as therapeutically acceptablesalts. The term “pharmaceuticallly acceptable salt,” as used herein,represents salts or zwitterionic forms of the compounds disclosed hereinwhich are therapeutically acceptable as defined herein. The salts can beprepared during the final isolation and purification of the compounds orseparately by reacting the appropriate compound with a suitable acid orbase. Therapeutically acceptable salts include acid and basic additionsalts. For a more complete discussion of the preparation and selectionof salts, refer to “Handbook of Pharmaceutical Salts, Properties, andUse,” Stah and Wermuth, Ed., (Wiley-VCH and VHCA, Zurich, 2002) andBerge et al., J. Pharm. Sci. 1977, 66, 1-19.

Suitable acids for use in the preparation of pharmaceutically acceptablesalts include, but are not limited to, acetic acid, 2,2-dichloroaceticacid, acylated amino acids, adipic acid, alginic acid, ascorbic acid,L-aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoicacid, boric acid, (+)-camphoric acid, camphorsulfonic acid,(+)-(1S)-camphor-10-sulfonic acid, capric acid, caproic acid, caprylicacid, cinnamic acid, citric acid, cyclamic acid, cyclohexanesulfamicacid, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonicacid, 2-hydroxy-ethanesulfonic acid, formic acid, fumaric acid,galactaric acid, gentisic acid, glucoheptonic acid, D-gluconic acid,D-glucuronic acid, L-glutamic acid, α-oxo-glutaric acid, glycolic acid,hippuric acid, hydrobromic acid, hydrochloric acid, hydroiodic acid,(+)-L-lactic acid, (±)-DL-lactic acid, lactobionic acid, lauric acid,maleic acid, (−)-L-malic acid, malonic acid, (±)-DL-mandelic acid,methanesulfonic acid, naphthalene-2-sulfonic acid,naphthalene-1,5-disulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinicacid, nitric acid, oleic acid, orotic acid, oxalic acid, palmitic acid,pamoic acid, perchloric acid, phosphoric acid, L-pyroglutamic acid,saccharic acid, salicylic acid, 4-amino-salicylic acid, sebacic acid,stearic acid, succinic acid, sulfuric acid, tannic acid, (+)-L-tartaricacid, thiocyanic acid, p-toluenesulfonic acid, undecylenic acid, andvaleric acid.

Suitable bases for use in the preparation of pharmaceutically acceptablesalts, including, but not limited to, inorganic bases, such as magnesiumhydroxide, calcium hydroxide, potassium hydroxide, zinc hydroxide, orsodium hydroxide; and organic bases, such as primary, secondary,tertiary, and quaternary, aliphatic and aromatic amines, includingL-arginine, benethamine, benzathine, choline, deanol, diethanolamine,diethylamine, dimethylamine, dipropylamine, diisopropylamine,2-(diethylamino)-ethanol, ethanolamine, ethylamine, ethylenediamine,isopropylamine, N-methyl-glucamine, hydrabamine, 1H-imidazole, L-lysine,morpholine, 4-(2-hydroxyethyl)-morpholine, methylamine, piperidine,piperazine, propylamine, pyrrolidine, 1-(2-hydroxyethyl)-pyrrolidine,pyridine, quinuclidine, quinoline, isoquinoline, secondary amines,triethanolamine, trimethylamine, triethylamine, N-methyl-D-glucamine,2-amino-2-(hydroxymethyl)-1,3-propanediol, and tromethamine.

While it may be possible for the compounds of the subject invention tobe administered as the raw chemical, it is also possible to present themas a pharmaceutical composition. Accordingly, provided herein arepharmaceutical compositions which comprise one or more of certaincompounds disclosed herein, or one or more pharmaceutically acceptablesalts, prodrugs, or solvates thereof, together with one or morepharmaceutically acceptable carriers thereof and optionally one or moreother therapeutic ingredients. Proper formulation is dependent upon theroute of administration chosen. Any of the well-known techniques,carriers, and excipients may be used as suitable and as understood inthe art; e.g., in Remington's Pharmaceutical Sciences. Thepharmaceutical compositions disclosed herein may be manufactured in anymanner known in the art, e.g., by means of conventional mixing,dissolving, granulating, dragee-making, levigating, emulsifying,encapsulating, entrapping or compression processes. The pharmaceuticalcompositions may also be formulated as a modified release dosage form,including delayed-, extended-, prolonged-, sustained-, pulsatile-,controlled-, accelerated- and fast-, targeted-, programmed-release, andgastric retention dosage forms. These dosage forms can be preparedaccording to conventional methods and techniques known to those skilledin the art (see, Remington: The Science and Practice of Pharmacy, supra;Modified-Release Drug Deliver Technology, Rathbone et al., Eds., Drugsand the Pharmaceutical Science, Marcel Dekker, Inc.: New York, N.Y.,2002; Vol. 126).

The compositions include those suitable for oral, parenteral (includingsubcutaneous, intradermal, intramuscular, intravenous, intraarticular,and intramedullary), intraperitoneal, transmucosal, transdermal, rectaland topical (including dermal, buccal, sublingual and intraocular)administration although the most suitable route may depend upon forexample the condition and disorder of the recipient. The compositionsmay conveniently be presented in unit dosage form and may be prepared byany of the methods well known in the art of pharmacy. Typically, thesemethods include the step of bringing into association a compound of thesubject invention or a pharmaceutically salt, prodrug, or solvatethereof (“active ingredient”) with the carrier which constitutes one ormore accessory ingredients. In general, the compositions are prepared byuniformly and intimately bringing into association the active ingredientwith liquid carriers or finely divided solid carriers or both and then,if necessary, shaping the product into the desired formulation.

Formulations of the compounds disclosed herein suitable for oraladministration may be presented as discrete units such as capsules,cachets or tablets each containing a predetermined amount of the activeingredient; as a powder or granules; as a solution or a suspension in anaqueous liquid or a non-aqueous liquid; or as an oil-in-water liquidemulsion or a water-in-oil liquid emulsion. The active ingredient mayalso be presented as a bolus, electuary or paste.

Pharmaceutical preparations which can be used orally include tablets,push-fit capsules made of gelatin, as well as soft, sealed capsules madeof gelatin and a plasticizer, such as glycerol or sorbitol. Tablets maybe made by compression or molding, optionally with one or more accessoryingredients. Compressed tablets may be prepared by compressing in asuitable machine the active ingredient in a free-flowing form such as apowder or granules, optionally mixed with binders, inert diluents, orlubricating, surface active or dispersing agents. Molded tablets may bemade by molding in a suitable machine a mixture of the powdered compoundmoistened with an inert liquid diluent. The tablets may optionally becoated or scored and may be formulated so as to provide slow orcontrolled release of the active ingredient therein. All formulationsfor oral administration should be in dosages suitable for suchadministration. The push-fit capsules can contain the active ingredientsin admixture with filler such as lactose, binders such as starches,and/or lubricants such as talc or magnesium stearate and, optionally,stabilizers. In soft capsules, the active compounds may be dissolved orsuspended in suitable liquids, such as fatty oils, liquid paraffin, orliquid polyethylene glycols. In addition, stabilizers may be added.Dragee cores are provided with suitable coatings. For this purpose,concentrated sugar solutions may be used, which may optionally containgum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethyleneglycol, and/or titanium dioxide, lacquer solutions, and suitable organicsolvents or solvent mixtures. Dyestuffs or pigments may be added to thetablets or dragee coatings for identification or to characterizedifferent combinations of active compound doses.

The compounds may be formulated for parenteral administration byinjection, e.g., by bolus injection or continuous infusion. Formulationsfor injection may be presented in unit dosage form, e.g., in ampoules orin multi-dose containers, with an added preservative. The compositionsmay take such forms as suspensions, solutions or emulsions in oily oraqueous vehicles, and may contain formulatory agents such as suspending,stabilizing and/or dispersing agents. The formulations may be presentedin unit-dose or multi-dose containers, for example sealed ampoules andvials, and may be stored in powder form or in a freeze-dried(lyophilized) condition requiring only the addition of the sterileliquid carrier, for example, saline or sterile pyrogen-free water,immediately prior to use. Extemporaneous injection solutions andsuspensions may be prepared from sterile powders, granules and tabletsof the kind previously described.

Formulations for parenteral administration include aqueous andnon-aqueous (oily) sterile injection solutions of the active compoundswhich may contain antioxidants, buffers, bacteriostats and solutes whichrender the formulation isotonic with the blood of the intendedrecipient; and aqueous and non-aqueous sterile suspensions which mayinclude suspending agents and thickening agents. Suitable lipophilicsolvents or vehicles include fatty oils such as sesame oil, or syntheticfatty acid esters, such as ethyl oleate or triglycerides, or liposomes.Aqueous injection suspensions may contain substances which increase theviscosity of the suspension, such as sodium carboxymethyl cellulose,sorbitol, or dextran. Optionally, the suspension may also containsuitable stabilizers or agents which increase the solubility of thecompounds to allow for the preparation of highly concentrated solutions.

In addition to the formulations described previously, the compounds mayalso be formulated as a depot preparation. Such long acting formulationsmay be administered by implantation (for example subcutaneously orintramuscularly) or by intramuscular injection. Thus, for example, thecompounds may be formulated with suitable polymeric or hydrophobicmaterials (for example as an emulsion in an acceptable oil) or ionexchange resins, or as sparingly soluble derivatives, for example, as asparingly soluble salt.

For buccal or sublingual administration, the compositions may take theform of tablets, lozenges, pastilles, or gels formulated in conventionalmanner. Such compositions may comprise the active ingredient in aflavored basis such as sucrose and acacia or tragacanth.

The compounds may also be formulated in rectal compositions such assuppositories or retention enemas, e.g., containing conventionalsuppository bases such as cocoa butter, polyethylene glycol, or otherglycerides.

Certain compounds disclosed herein may be administered topically, thatis by non-systemic administration. This includes the application of acompound disclosed herein externally to the epidermis or the buccalcavity and the instillation of such a compound into the ear, eye andnose, such that the compound does not significantly enter the bloodstream. In contrast, systemic administration refers to oral,intravenous, intraperitoneal and intramuscular administration.

Formulations suitable for topical administration include liquid orsemi-liquid preparations suitable for penetration through the skin tothe site of inflammation such as gels, liniments, lotions, creams,ointments or pastes, and drops suitable for administration to the eye,ear or nose.

For administration by inhalation, compounds may be delivered from aninsufflator, nebulizer pressurized packs or other convenient means ofdelivering an aerosol spray. Pressurized packs may comprise a suitablepropellant such as dichlorodifluoromethane, trichlorofluoromethane,dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In thecase of a pressurized aerosol, the dosage unit may be determined byproviding a valve to deliver a metered amount. Alternatively, foradministration by inhalation or insufflation, the compounds according tothe invention may take the form of a dry powder composition, for examplea powder mix of the compound and a suitable powder base such as lactoseor starch. The powder composition may be presented in unit dosage form,in for example, capsules, cartridges, gelatin or blister packs fromwhich the powder may be administered with the aid of an inhalator orinsufflator.

Preferred unit dosage formulations are those containing an effectivedose, as herein below recited, or an appropriate fraction thereof, ofthe active ingredient.

Compounds may be administered orally or via injection at a dose of from0.1 to 500 mg/kg per day. The dose range for adult humans is generallyfrom 5 mg to 2 g/day. Tablets or other forms of presentation provided indiscrete units may conveniently contain an amount of one or morecompounds which is effective at such dosage or as a multiple of thesame, for instance, units containing 5 mg to 500 mg, usually around 10mg to 200 mg.

The amount of active ingredient that may be combined with the carriermaterials to produce a single dosage form will vary depending upon thehost treated and the particular mode of administration.

The compounds can be administered in various modes, e.g. orally,topically, or by injection. The precise amount of compound administeredto a patient will be the responsibility of the attendant physician. Thespecific dose level for any particular patient will depend upon avariety of factors including the activity of the specific compoundemployed, the age, body weight, general health, sex, diets, time ofadministration, route of administration, rate of excretion, drugcombination, the precise disorder being treated, and the severity of thedisorder being treated. Also, the route of administration may varydepending on the disorder and its severity.

In the case wherein the patient's condition does not improve, upon thedoctor's discretion the administration of the compounds may beadministered chronically, that is, for an extended period of time,including throughout the duration of the patient's life in order toameliorate or otherwise control or limit the symptoms of the patient'sdisorder.

In the case wherein the patient's status does improve, upon the doctor'sdiscretion the administration of the compounds may be given continuouslyor temporarily suspended for a certain length of time (i.e., a “drugholiday”).

Once improvement of the patient's conditions has occurred, a maintenancedose is administered if necessary. Subsequently, the dosage or thefrequency of administration, or both, can be reduced, as a function ofthe symptoms, to a level at which the improved disorder is retained.Patients can, however, require intermittent treatment on a long-termbasis upon any recurrence of symptoms.

Disclosed herein are methods of treating a P2Y12 receptor-mediateddisorder comprising administering to a subject having or suspected ofhaving such a disorder, a therapeutically effective amount of a compoundas disclosed herein or a pharmaceutically acceptable salt, solvate, orprodrug thereof.

P2Y12 receptor-mediated disorders, include, but are not limited to,arterial thrombosis, coronary artery disease, myocardial infarction,stroke, atherosclerosis, acute coronary syndrome, peripheral arteryocclusive disease, carotid, vertebral, or intracerebral artery stenosis,unstable angina, primary arterial thrombotic complications ofatherosclerosis such as thrombotic or embolic stroke, transientischaemic attacks, peripheral vascular disease, myocardial infarctionwith or without thrombolysis, arterial complications due tointerventions in atherosclerotic disease such as angioplasty, includingcoronary angioplasty (PTCA), endarterectomy, stent placement, coronaryand other vascular graft surgery, thrombotic complications of surgicalor mechanical damage such as tissue salvage following accidental orsurgical trauma, reconstructive surgery including skin and muscle flaps,conditions with a diffuse thrombotic/platelet consumption component suchas disseminated intravascular coagulation, thrombotic thrombocytopaenicpurpura, haemolytic uraemic syndrome, thrombotic complications ofsepticaemia, adult respiratory distress syndrome, anti-phospholipidsyndrome, heparin-induced thrombocytopaenia and pre-eclampsia/eclampsia,or venous thrombosis such as deep vein thrombosis, venoocclusivedisease, haematological conditions such as myeloproliferative disease,including thrombocythaemia, sickle cell disease; or in the prevention ofmechanically-induced platelet activation in vivo, such ascardio-pulmonary bypass and extracorporeal membrane oxygenation(prevention of microthromboembolism), mechanically-induced plateletactivation in vitro, such as use in the preservation of blood products,e.g. platelet concentrates, or shunt occlusion such as in renal dialysisand plasmapheresis, thrombosis secondary to vascular damage/inflammationsuch as vasculitis, arteritis, glomerulonephritis, inflammatory boweldisease and organ graft rejection, conditions such as migraine,Raynaud's phenomenon, conditions in which platelets can contribute tothe underlying inflammatory disease process in the vascular wall such asatheromatous plaque formation/progression, stenosis/restenosis, in otherinflammatory conditions such as asthma, in which platelets andplatelet-derived factors are implicated in the immunological diseaseprocess, and/or any disorder which can lessened, alleviated, orprevented by administering a P2Y12 receptor modulator.

In certain embodiments, a method of treating a P2Y12 receptor-mediateddisorder comprises administering to the subject a therapeuticallyeffective amount of a compound as disclosed herein, or apharmaceutically acceptable salt, solvate, or prodrug thereof, so as toaffect: (1) decreased inter-individual variation in plasma levels of thecompound or a metabolite thereof; (2) increased average plasma levels ofthe compound or decreased average plasma levels of at least onemetabolite of the compound per dosage unit; (3) decreased inhibition of,and/or metabolism by at least one cytochrome P₄₅₀ or monoamine oxidaseisoform in the subject; (4) decreased metabolism via at least onepolymorphically-expressed cytochrome P₄₅₀ isoform in the subject; (5) atleast one statistically-significantly improved disorder-control and/ordisorder-eradication endpoint; (6) an improved clinical effect duringthe treatment of the disorder, (7) prevention of recurrence, or delay ofdecline or appearance, of abnormal alimentary or hepatic parameters asthe primary clinical benefit, or (8) reduction or elimination ofdeleterious changes in any diagnostic hepatobiliary function endpoints,as compared to the corresponding non-isotopically enriched compound.

In certain embodiments, inter-individual variation in plasma levels ofthe compounds as disclosed herein, or metabolites thereof, is decreased;average plasma levels of the compound as disclosed herein are increased;average plasma levels of a metabolite of the compound as disclosedherein are decreased; inhibition of a cytochrome P₄₅₀ or monoamineoxidase isoform by a compound as disclosed herein is decreased; ormetabolism of the compound as disclosed herein by at least onepolymorphically-expressed cytochrome P₄₅₀ isoform is decreased; bygreater than about 5%, greater than about 10%, greater than about 20%,greater than about 30%, greater than about 40%, or by greater than about50% as compared to the corresponding non-isotopically enriched compound.

Plasma levels of the compound as disclosed herein, or metabolitesthereof, may be measured using the methods described by Li et al. RapidCommunications in Mass Spectrometry 2005, 19, 1943-1950; Butler, et al.,Drug Metab Rev 2008, 40(Suppl. 3): Abst 280; Husted et al., EuropeanHeart Journal 2006, 27(9), 1038-1047; and any references cited thereinand any modifications made thereof.

Examples of cytochrome P₄₅₀ isoforms in a mammalian subject include, butare not limited to, CYP1A1, CYP1A2, CYP1B1, CYP2A6, CYP2A13, CYP2B6,CYP2C8, CYP2C9, CYP2C18, CYP2C19, CYP2D6, CYP2E1, CYP2G1, CYP2J2,CYP2R1, CYP2S1, CYP3A4, CYP3A5, CYP3A5P1, CYP3A5P2, CYP3A7, CYP4A11,CYP4B1, CYP4F2, CYP4F3, CYP4F8, CYP4F11, CYP4F12, CYP4X1, CYP4Z1,CYP5A1, CYP7A1, CYP7B1, CYP8A1, CYP8B1, CYP11A1, CYP11B1, CYP11B2,CYP17, CYP19, CYP21, CYP24, CYP26A1, CYP26B1, CYP27A1, CYP27B1, CYP39,CYP46, and CYP51.

Examples of monoamine oxidase isoforms in a mammalian subject include,but are not limited to, MAO_(A), and MAO_(B).

The inhibition of the cytochrome P₄₅₀ isoform is measured by the methodof Ko et al., British Journal of Clinical Pharmacology 2000, 49,343-351. The inhibition of the MAO_(A) isoform is measured by the methodof Weyler et al., J. Biol. Chem. 1985, 260, 13199-13207. The inhibitionof the MAO_(B) isoform is measured by the method of Uebelhack et al.,Pharmacopsychiatry, 1998, 31, 187-192.

Examples of polymorphically-expressed cytochrome P₄₅₀ isoforms in amammalian subject include, but are not limited to, CYP2C8, CYP2C9,CYP2C19, and CYP2D6.

The metabolic activities of liver microsomes, cytochrome P₄₅₀ isoforms,and monoamine oxidase isoforms are measured by the methods describedherein.

Examples of improved disorder-control and/or disorder-eradicationendpoints, or improved clinical effects include, but are not limited to,bleeding time, platelet inhibition, inhibition ofadenosine-5′-diphosphate-induced platelet aggregation as measured byoptical aggregometry of platelet-rich plasma, reduced cardiovasculardeath, reduced myocardial infarction, reduced stroke, and reducedbleeding events (Tantry et al., Exp. Opin. Invest. Drugs 2007, 16(2),225-229; Husted et al., Eur. Heart J. 2006, 27(9), 1038-1047; and WO2000034283).

Examples of diagnostic hepatobiliary function endpoints include, but arenot limited to, alanine aminotransferase (“ALT”), serum glutamic-pyruvictransaminase (“SGPT”), aspartate aminotransferase (“AST” or “SGOT”),ALT/AST ratios, serum aldolase, alkaline phosphatase (“ALP”), ammonialevels, bilirubin, gamma-glutamyl transpeptidase (“GGTP,” “γ-GTP,” or“GGT”), leucine aminopeptidase (“LAP”), liver biopsy, liverultrasonography, liver nuclear scan, 5′-nucleotidase, and blood protein.Hepatobiliary endpoints are compared to the stated normal levels asgiven in “Diagnostic and Laboratory Test Reference”, 4^(th) edition,Mosby, 1999. These assays are run by accredited laboratories accordingto standard protocol.

Besides being useful for human treatment, certain compounds andformulations disclosed herein may also be useful for veterinarytreatment of companion animals, exotic animals and farm animals,including mammals, rodents, and the like. More preferred animals includehorses, dogs, and cats.

Combination Therapy

The compounds disclosed herein may also be combined or used incombination with other agents useful in the treatment of P2Y12receptor-mediated disorders. Or, by way of example only, the therapeuticeffectiveness of one of the compounds described herein may be enhancedby administration of an adjuvant (i.e., by itself the adjuvant may onlyhave minimal therapeutic benefit, but in combination with anothertherapeutic agent, the overall therapeutic benefit to the patient isenhanced).

Such other agents, adjuvants, or drugs, may be administered, by a routeand in an amount commonly used therefor, simultaneously or sequentiallywith a compound as disclosed herein. When a compound as disclosed hereinis used contemporaneously with one or more other drugs, a pharmaceuticalcomposition containing such other drugs in addition to the compounddisclosed herein may be utilized, but is not required.

In certain embodiments, the compounds disclosed herein can be combinedwith one or more alpha adrenergic receptor antagonists, beta-adrenergicantagonists, angiotensin II receptor antagonists, angiotensin-convertingenzyme inhibitors, anti-arrhythmics, antithrombotics, antiplateletagents, calcium channel blockers, fibrates, and HMG-CoA reductaseinhibitors.

In certain embodiments, the compounds disclosed herein can be combinedwith one or more alpha adrenergic receptor antagonists known in the art,including, but not limited to, abanoquil, adimolol, ajmalicine,alfuzosin, amosulalol, arotinolol, atiprosin, benoxathian, buflomedil,bunazosin, carvedilol, CI-926, corynanthine, dapiprazole, DL-017,domesticine, doxazosin, eugenodilol, fenspiride, GYKI-12,743,GYKI-16,084, indoramin, ketanserin, L-765,314, labetalol, mephendioxan,metazosin, monatepil, moxisylyte (thymoxamine), naftopidil, nantenine,neldazosin, nicergoline, niguldipine, pelanserin, phendioxan,phenoxybenzamine, phentolamine, piperoxan, prazosin, quinazosin,ritanserin, RS-97,078, SGB-1,534, silodosin, SL-89.0591, spiperone,talipexole, tamsulosin, terazosin, tibalosin, tiodazosin, tipentosin,tolazoline, trimazosin, upidosin, urapidil, zolertine, 1-PP, adimolol,atipamezole, BRL-44408, buflomedil, cirazoline, efaroxan, esmirtazapine,fluparoxan, GYKI-12,743, GYKI-16,084, idazoxan, mianserin, mirtazapine,MK-912, NAN-190, olanzapine, phentolamine, phenoxybenzamine, piperoxan,piribedil, rauwolscine, rotigotine, SB-269,970, setiptiline,spiroxatrine, sunepitron, tolazoline, and yohimbine.

In certain embodiments, the compounds disclosed herein can be combinedwith one or more beta-adrenergic antagonists, including, but not limitedto, acebutolol, adaprolol, adimolol, afurolol, alprenolol, alprenoxime,amosulalol, ancarolol, arnolol, arotinolol, atenolol, befunolol,betaxolol, bevantolol, bisoprolol, bopindolol, bormetolol, bornaprolol,brefonalol, bucindolol, bucumolol, bufetolol, buftiralol, bufuralol,bunitrolol, bunolol, bupranolol, burocrolol, butaxamine, butidrine,butofilolol, capsinolol, carazolol, carpindolol, carteolol, carvedilol,celiprolol, cetamolol, cicloprolol, cinamolol, cloranolol,cyanopindolol, dalbraminol, dexpropranolol, diacetolol,dichloroisoprenaline, dihydroalprenolol, dilevalol, diprafenone,draquinolol, dropranolol, ecastolol, epanolol, ericolol, ersentilide,esatenolol, esmolol, esprolol, eugenodilol, exaprolol, falintolol,flestolol, flusoxolol, hydroxycarteolol, hydroxytertatolol, ICI-118,551,idropranolol, indenolol, indopanolol, iodocyanopindolol, iprocrolol,isoxaprolol, isamoltane, labetalol, landiolol, levobetaxolol,levobunolol, levocicloprolol, levomoprolol, medroxalol, mepindolol,metalol, metipranolol, metoprolol, moprolol, nadolol, nadoxolol,nafetolol, nebivolol, neraminol, nifenalol, nipradilol, oberadilol,oxprenolol, pacrinolol, pafenolol, pamatolol, pargolol, parodilol,penbutolol, penirolol, PhQA-33, pindolol, pirepolol, practolol,primidolol, procinolol, pronethalol, propafenone, propranolol,ridazolol, ronactolol, soquinolol, sotalol, spirendolol, SR 59230A,sulfinalol, TA-2005, talinolol, tazolol, teoprolol, tertatolol,terthianolol, tienoxolol, tilisolol, timolol, tiprenolol, tolamolol,toliprolol, tribendilol, trigevolol, xibenolol, and xipranolol.

In certain embodiments, the compounds disclosed herein can be combinedwith one or more angiotensin II receptor antagonists, including, but notlimited to, candesartan, eprosartan, irbesartan, losartan, olmesartan,tasosartan, telmisartan, and valsartan.

In certain embodiments, the compounds disclosed herein can be combinedwith one or more angiotensin-converting enzyme inhibitors, including,but not limited to, captopril, enalapril, lisinopril, perindopril,ramipril, quinapril, benazepril, cilazapril, fosinopril, trandolapril,spirapril, delapril, moexipril, temocapril, zofenopril, and imidapril.

In certain embodiments, the compounds disclosed herein can be combinedwith one or more anti-arrhythmics, including, but not limited toquinidine, procainamide, disopyramide, sparteine, ajmaline, prajmaline,lorajmine, lidocaine, mexiletine, tocamide, aprindine, propafenone,flecamide, lorcamide, encamide, amiodarone, bretylium tosilate,bunaftine, dofetilide, ibutilidem, tedisamil, moracizine, andcibenzoline.

In certain embodiments, the compounds provided herein can be combinedwith one or more antithrombotics, including, but not limited to,dicoumarol, phenindione, warfarin, phenprocoumon, acenocoumarol, ethylbiscoumacetate, clorindione, diphenadione, tioclomarol, heparin,antithrombin III, dalteparin, enoxaparin, nadroparin, parnaparin,reviparin, danaparoid, tinzaparin, sulodexide, bemiparin, ditazole,cloricromen, picotamide, clopidogrel, ticlopidine, acetylsalicylic acid,dipyridamole, carbasalate calcium, epoprostenol, indobufen, iloprost,abciximab, aloxiprin, eptifibatide, tirofiban, triflusal, beraprost,treprostinil, prasugrel, streptokinase, alteplase, urokinase,fibrinolysin, brinase, reteplase, saruplase, ancrod, drotrecogin alfa(activated), tenecteplase, protein C, desirudin, lepirudin, argatroban,melagatran, ximelagatran, bivalirudin, dabigatran etexilate,defibrotide, dermatan sulfate, fondaparinux, and rivaroxaban.

In certain embodiments, the compounds provided herein can be combinedwith one or more antiplatelet agents, including, but not limited to,abciximab, eptifibatide, tirofiban, clopidogrel, prasugrel, ticlopidine,ticagrelor, beraprost, prostacyclin, iloprost, treprostinil,acetylsalicylic acid, aloxiprin, carbasalate calcium, indobufen,dipyridamole, picotamide, terutroban, cilostazol, dipyridamole,triflusal, cloricromen, and ditazole.

In certain embodiments, the compounds disclosed herein can be combinedwith one or more beta-adrenergic antagonists, including, but not limitedto, acebutolol, adaprolol, adimolol, afurolol, alprenolol, alprenoxime,amosulalol, ancarolol, arnolol, arotinolol, atenolol, befunolol,betaxolol, bevantolol, bisoprolol, bopindolol, bormetolol, bornaprolol,brefonalol, bucindolol, bucumolol, bufetolol, buftiralol, bufuralol,bunitrolol, bunolol, bupranolol, burocrolol, butaxamine, butidrine,butofilolol, capsinolol, carazolol, carpindolol, carteolol, carvedilol,celiprolol, cetamolol, cicloprolol, cinamolol, cloranolol,cyanopindolol, dalbraminol, dexpropranolol, diacetolol,dichloroisoprenaline, dihydroalprenolol, dilevalol, diprafenone,draquinolol, dropranolol, ecastolol, epanolol, ericolol, ersentilide,esatenolol, esmolol, esprolol, eugenodilol, exaprolol, falintolol,flestolol, flusoxolol, hydroxycarteolol, hydroxytertatolol, ICI-118,551,idropranolol, indenolol, indopanolol, iodocyanopindolol, iprocrolol,isoxaprolol, isamoltane, labetalol, landiolol, levobetaxolol,levobunolol, levocicloprolol, levomoprolol, medroxalol, mepindolol,metalol, metipranolol, metoprolol, moprolol, nadolol, nadoxolol,nafetolol, nebivolol, neraminol, nifenalol, nipradilol, oberadilol,oxprenolol, pacrinolol, pafenolol, pamatolol, pargolol, parodilol,penbutolol, penirolol, PhQA-33, pindolol, pirepolol, practolol,primidolol, procinolol, pronethalol, propafenone, propranolol,ridazolol, ronactolol, soquinolol, sotalol, spirendolol, SR 59230A,sulfinalol, TA-2005, talinolol, tazolol, teoprolol, tertatolol,terthianolol, tienoxolol, tilisolol, timolol, tiprenolol, tolamolol,toliprolol, tribendilol, trigevolol, xibenolol, and xipranolol.

In certain embodiments, the compounds disclosed herein can be combinedwith one or more calcium channel blockers, including, but not limited toamlodipine, felodipine, isradipine, nicardipine, nifedipine, nimodipine,nisoldipine, nitrendipine, lacidipine, nilvadipine, manidipine,barnidipine, lercanidipine, cilnidipine, benidipine, mibefradil,verapamil, gallopamil, diltiazem, fendiline, bepridil, lidoflazine, andperhexyline.

In certain embodiments, the compounds provided herein can be combinedwith one or more fibrates, including, but not limited to, clofibrate,bezafibrate, aluminium clofibrate, gemfibrozil, fenofibrate, simfibrate,ronifibrate, ciprofibrate, etofibrate, and clofibride.

In certain embodiments, the compounds disclosed herein can be combinedwith one or more HMG-CoA reductase inhibitors, including, but notlimited to, atorvastatin, cerivastatin, fluvastatin, lovastatin,mevastatin, pitavastatin, pravastatin, rosuvastatin, and simvastatin.

The compounds disclosed herein can also be administered in combinationwith other classes of compounds, including, but not limited to,decongestant treatments; antitussive treatments; mucolytic treatments;expectorant treatments; antiallergic non-steroidal treatments; steroidaldrugs; antihistamine treatments; leukotriene receptor antagonists;phosphodiesterase inhibitors; CYP3A inhibitors; CYP3A inducers; proteaseinhibitors; antifugal agents; antibacterials; antimycobacterial agents;sepsis treatments; steroidal drugs; non-steroidal anti-inflammatoryagents, norepinephrine reuptake inhibitors (NRIs) such as atomoxetine;dopamine reuptake inhibitors (DARIs), such as methylphenidate;serotonin-norepinephrine reuptake inhibitors (SNRIs), such asmilnacipran; sedatives, such as diazepham; norepinephrine-dopaminereuptake inhibitor (NDRIs), such as bupropion;serotonin-norepinephrine-dopamine-reuptake-inhibitors (SNDRIs), such asvenlafaxine; monoamine oxidase inhibitors, such as selegiline;hypothalamic phospholipids; endothelin converting enzyme (ECE)inhibitors, such as phosphoramidon; opioids, such as tramadol;thromboxane receptor antagonists, such as ifetroban; potassium channelopeners; thrombin inhibitors, such as hirudin; hypothalamicphospholipids; growth factor inhibitors, such as modulators of PDGFactivity; platelet activating factor (PAF) antagonists; Factor VIIaInhibitors and Factor Xa Inhibitors; renin inhibitors; neutralendopeptidase (NEP) inhibitors; vasopepsidase inhibitors (dual NEP-ACEinhibitors), such as omapatrilat and gemopatrilat; squalene synthetaseinhibitors; bile acid sequestrants, such as questran; niacin;anti-atherosclerotic agents, such as ACAT inhibitors; MTP Inhibitors;potassium channel activators; alpha-muscarinic agents; beta-muscarinicagents, such as carvedilol and metoprolol; diuretics, such aschlorothlazide, hydrochlorothiazide, flumethiazide, hydroflumethiazide,bendroflumethiazide, methylchlorothiazide, trichloromethiazide,polythiazide, benzothlazide, ethacrynic acid, tricrynafen,chlorthalidone, furosenilde, musolimine, bumetanide, triamterene,amiloride, and spironolactone; anti-diabetic agents, such as biguanides(e.g. metformin), glucosidase inhibitors (e.g., acarbose), insulins,meglitinides (e.g., repaglinide), sulfonylureas (e.g., glimepiride,glyburide, and glipizide), thiozolidinediones (e.g. troglitazone,rosiglitazone and pioglitazone), and PPAR-gamma agonists;mineralocorticoid receptor antagonists, such as spironolactone andeplerenone; growth hormone secretagogues; aP2 inhibitors;phosphodiesterase inhibitors, such as PDE III inhibitors (e.g.,cilostazol) and PDE V inhibitors (e.g., sildenafil, tadalafil,vardenafil); protein tyrosine kinase inhibitors; antiinflammatories;antiproliferatives, such as methotrexate, FK506 (tacrolimus, Prograf),mycophenolate mofetil; chemotherapeutic agents; immunosuppressants;anticancer agents and cytotoxic agents (e.g., alkylating agents, such asnitrogen mustards, alkyl sulfonates, nitrosoureas, ethylenimines, andtriazenes); antimetabolites, such as folate antagonists, purineanalogues, and pyrridine analogues; antibiotics, such as anthracyclines,bleomycins, mitomycin, dactinomycin, and plicamycin; enzymes, such asL-asparaginase; farnesyl-protein transferase inhibitors; hormonalagents, such as glucocorticoids (e.g., cortisone),estrogens/antiestrogens, androgens/antiandrogens, progestins, andluteinizing hormone-releasing hormone anatagonists, and octreotideacetate; microtubule-disruptor agents, such as ecteinascidins;microtubule-stablizing agents, such as pacitaxel, docetaxel, andepothilones A-F; plant-derived products, such as vinca alkaloids,epipodophyllotoxins, and taxanes; and topoisomerase inhibitors;prenyl-protein transferase inhibitors; and cyclosporins; steroids, suchas prednisone and dexamethasone; cytotoxic drugs, such as azathiprineand cyclophosphamide; TNF-alpha inhibitors, such as tenidap; anti-TNFantibodies or soluble TNF receptor, such as etanercept, rapamycin, andleflunimide; and cyclooxygenase-2 (COX-2) inhibitors, such as celecoxiband rofecoxib; and miscellaneous agents such as, hydroxyurea,procarbazine, mitotane, hexamethylmelamine, gold compounds, platinumcoordination complexes, such as cisplatin, satraplatin, and carboplatin.

Thus, in another aspect, certain embodiments provide methods fortreating P2Y12 receptor-mediated disorders in a human or animal subjectin need of such treatment comprising administering to said subject anamount of a compound disclosed herein effective to reduce or preventsaid disorder in the subject, in combination with at least oneadditional agent for the treatment of said disorder that is known in theart. In a related aspect, certain embodiments provide therapeuticcompositions comprising at least one compound disclosed herein incombination with one or more additional agents for the treatment ofP2Y12 receptor-mediated disorders.

General Synthetic Methods for Preparing Compounds

Isotopic hydrogen can be introduced into a compound as disclosed hereinby synthetic techniques that employ deuterated reagents, wherebyincorporation rates are predetermined; and/or by exchange techniques,wherein incorporation rates are determined by equilibrium conditions,and may be highly variable depending on the reaction conditions.Synthetic techniques, where tritium or deuterium is directly andspecifically inserted by tritiated or deuterated reagents of knownisotopic content, may yield high tritium or deuterium abundance, but canbe limited by the chemistry required. Exchange techniques, on the otherhand, may yield lower tritium or deuterium incorporation, often with theisotope being distributed over many sites on the molecule.

The compounds as disclosed herein can be prepared by methods known toone of skill in the art and routine modifications thereof, and/orfollowing procedures similar to those described in the Example sectionherein and routine modifications thereof, and/or procedures found inShireman et al., Tetrahedron Letters 2000, 41, 9537-9540; Bioorganic &Medicinal Chemistry Letters 2007, 17, 6013-6018; US 20030148888 and WO2010030224; WO 2000034283; WO 2001092262; WO 2001092263; WO 199905142,which are hereby incorporated in their entirety, and references citedtherein and routine modifications thereof. Compounds as disclosed hereincan also be prepared as shown in any of the following schemes androutine modifications thereof.

The following schemes can be used to practice the present invention. Anyposition shown as hydrogen may optionally be replaced with deuterium.

Compound 1 is treated with malonic acid in the presence of anappropriate base, such as piperidine, in an appropriate solvent, such aspyridine, to give compound 2. Compound 2 is reacted with an appropriatechlorinating agent, such as thionyl chloride, in the presence of anappropriate base, such as pyridine, in an appropriate solvent, such astoluene, to give compound 3. Compound 3 is reacted with compound 4 inthe presence of an appropriate base, such as pyridine, in an appropriatesolvent, such as toluene, to give compound 5. Compound 5 is reacted withcompound 6, in the presence of an appropriate base, such as sodiumhydroxide, in an appropriate solvent, such as an appropriate mixture ofdimethyl sulfoxide and water, to give compound 7. Compound 7 is reactedwith an appropriate hydroxide base, such as sodium hydroxide, in anappropriate solvent, such as an appropriate mixture of dimethylsulfoxide and water, to give compound 8. Compound 8 is reacted with anappropriate chlorinating agent, such as thionyl chloride, in anappropriate solvent, such as toluene, to give an acyl cholideintermediate which is then reacted with an appropriate azide source,such as sodium azide, in the presence of an appropriate base, such assodium carbonate, in the presence of an appropriate phase-transfercatalyst, such as tetrabutylammonium bromide, to give compound 9.Compound 9 is reacted at an elevated temperature in an appropriatesolvent, such as toluene, to give compound 10. Compound 11 is reactedwith an appropriate amine protecting reagent, such as benzylchloroformate, in the presence of an appropriate base, such as potassiumcarbonate, in an appropriate solvent, such as 4-methyl-2-pentanone, togive compound 12. Compound 12 is reacted with compound 13 in thepresence of an appropriate base, such as potassium tert-butoxide, in anappropriate solvent, such as tetrahydrofuran, to give compound 14.Compound 14 is treated with an appropriate reducing reagent, such aslithium borohydride, in an appropriate solvent, such as tetrahydrofuran,to give compound 15. Compound 15 is treated with an appropriatedeprotecting reagent, such as a combination of hydrogen gas andpalladium on carbon, in an appropriate solvent, such as ethanol, to givecompound 16. Compound 17 is reacted with compound 18 in the presence ofan appropriate base, such as sodium hydroxide, in an appropriatesolvent, such as an appropriate mixture of water and1-methyl-2-pyrrolidinone, to give compound 19. Compound 19 is reactedwith an appropriate base, such as sodium hydroxide, and then reactedwith an aromatic amine, such as para-toluidine, in the presence of anappropriate nitrite salt, such as sodium nitrite, in the presence of anappropriate acid, such as hydrochloric acid, in an appropriate solvent,such as water, to give compound 20. Compound 20 is reacted with anappropriate chlorinating agent, such as phosphorous oxychloride, in anappropriate solvent, such as toluene, to give compound 21. Compound 21is reacted with an appropriate reducing agent, such as a hydrogen gas,in the presence of an appropriate catalyst, such as platinum on carbon,in an appropriate solvent, such as 2-propanol, to give compound 22.Compound 22 is reacted with compound 16, in the presence of anappropriate base, such as triethylamine, in an appropriate solvent, suchas ethanol, to give compound 23. Compound 23 is reacted with anappropriate nitrite salt, such as sodium nitrite, in the presence of anappropriate acid, such as acetic acid, in an appropriate solvent, suchas water, to give compound 24. Compound 24 is reacted with compound 10in the presence of an appropriate base, such as triethylamine, in anappropriate solvent, such as acetonitrile, to give compound 25. Compound25 is treated with an appropriate ketal deprotecting reagent, such ashydrochloric acid, in an appropriate solvent, such as a combination ofwater and methanol, to give a compound 26 of Formula I.

Deuterium can be incorporated to different positions synthetically,according to the synthetic procedures as shown in Scheme I, by usingappropriate deuterated intermediates. For example, to introducedeuterium at one or more positions of R₁-R₄, compound 1 with thecorresponding deuterium substitutions can be used. To introducedeuterium at R₇, malonic acid with the corresponding deuteriumsubstitutions can be used. To introduce deuterium at one or morepositions of R₅-R₆, compound 6 with the corresponding deuteriumsubstitutions can be used. To introduce deuterium at one or morepositions of R₁₆-R₂₁, compound 11 with the corresponding deuteriumsubstitutions can be used. To introduce deuterium at one or morepositions of R₂₄-R₂₅, compound 13 with the corresponding deuteriumsubstitutions can be used. To introduce deuterium at one or morepositions of R₂₆-R₂₇, lithium borodeuteride can be used. To introducedeuterium at one or more positions of R₉-R₁₅, compound 18 with thecorresponding deuterium substitutions can be used.

Deuterium can be incorporated to various positions having anexchangeable proton, such as the amine N—H and hydroxyl O—Hs, viaproton-deuterium equilibrium exchange. For example, to introducedeuterium at R₈, R₂₂-R₂₃, and R₂₈, this proton may be replaced withdeuterium selectively or non-selectively through a proton-deuteriumexchange method known in the art.

Compound 27 is reacted with compound 28 in the presence of anappropriate oxidant, such as a combination of oxalyl chloride, dimethylsulfoxide, and triethylamine, in an appropriate solvent, such as acombination of dichloromethane and dimethyl sulfoxide, to give compound29. Compound 29 is reacted with an appropriate oxidizing agent, such asa combination of osmium tetroxide and N-methylmorpholine N-oxide, in anappropriate solvent, such as a combination of water and tetrahydrofuran,to give compound 30. Compound 30 is reacted with an appropriate1,2-dihydroxy protecting group, such as 2,2-dimethoxypropane, in thepresence of an appropriate acid, such as p-toluenesulfonic acid, to givecompound 31. Compound 31 is reacted with an appropriate reducing agent,such as sodium borohydride, in an appropriate solvent, such as methanol,to give compound 32. Compound 32 is treated with an appropriate reducingagent, such as a combination of hydrogen gas and palladium on carbon, inan appropriate solvent, such as methanol, to give compound 33. Compound33 is reacted with an appropriate amine protecting reagent, such asbenzyl chloroformate, in the presence of an appropriate base, such aspotassium carbonate, in an appropriate solvent, such as a combination ofwater and tetrahydrofuran, to give compound 34. Compound 35 is treatedwith an appropriate reducing reagent, such as lithium aluminum hydride,in an appropriate solvent, such as tetrahydrofuran, to give compound 36.Compound 36 is reacted with an appropriate alcohol protecting reagent,such as benzyl bromide, in the presence of an appropriate base, such assilver oxide, in an appropriate solvent, such as dichloromethane, togive compound 37. Compound 37 is reacted with an appropriate brominatingagent, such as a combination of N-bromosuccinimide andtriphenylphosphine, in an appropriate solvent, such as tetrahydrofuran,to give compound 38. Compound 34 is reacted with compound 38 in thepresence of an appropriate base, such as sodium hydride, in anappropriate solvent, such as dimethylformamide, to give compound 39.Compound 39 is treated with an appropriate deprotecting reagent, such asa combination of hydrogen gas and palladium on carbon, in an appropriatesolvent, such as methanol, to give compound 16. Compound 40 is reactedwith compound 41 in the presence of an appropriate base, such as sodiumethoxide, in an appropriate solvent, such as ethanol, to give compound42. Compound 42 is reacted with an appropriate base, such astriethylamine, at an elevated temperature, in an appropriate solvent,such as methanol or d₄-methanol, to give compound 43. Compound 43 isreacted with an appropriate decarboxylating catalyst, such as sodiumchloride, in an appropriate solvent, such as a combination of water anddimethylsulfoxide, to give compound 44. Compound 44 is treated with anappropriate reducing reagent, such as lithium aluminum hydride, in anappropriate solvent, such as diethyl ether, to give compound 45.Compound 45 is treated with an appropriate iodinating reagent, such ashydroiodic acid, to give compound 18. Compound 18 is reacted withcompound 17 in the presence of an appropriate base, such as sodiumhydroxide, in an appropriate solvent, such as an appropriate mixture ofwater and 1-methyl-2-pyrrolidinone, to give compound 19. Compound 19 isreacted with an appropriate nitrating agent, such as nitric acid, togive compound 46. Compound 46 is reacted with an appropriatechlorinating agent, such as phosphorous oxychloride, in the presence ofan appropriate base, such as N,N-diethylbenzenamine, to give compound47. Compound 47 is reacted with compound 16 in an appropriate solvent,such as tetrahydrofuran, to give compound 48. Compound 48 is treatedwith an appropriate reducing reagent, such as a combination of iron andacetic acid, in an appropriate solvent, such as a mixture of ethanol andwater, to give compound 23. Compound 23 is reacted with an appropriatenitrite salt, such as sodium nitrite, in the presence of an appropriateacid, such as acetic acid, in an appropriate solvent, such as acombination of toluene and water, to give compound 24. Compound 2 isreacted with an appropriate chlorinating agent, such as oxalyl chloride,in the presence of an appropriate catalyst, such as dimethylformamide,in an appropriate solvent, such as dichloromethane, to give compound 3.Compound 3 is reacted with an appropriate chiral auxiliary, such as(2R)-bornane-10,2-sultam, in the presence of an appropriate base, suchas triethylamine, in an appropriate solvent, such as dichloromethane, togive compound 49. Compound 49 is reacted with compound 50, in thepresence of an appropriate catalyst, such as palladium (II) acetate, inan appropriate solvent, such as an appropriate mixture of diethyl etherand dichloromethane, to give compound 51. Compound 51 is reacted with anappropriate base, such as lithium hydroxide, in an appropriate solvent,such as an appropriate mixture of tetrahydrofuran and water, to givecompound 8. Compound 8 is reacted with an appropriate acyl azide-formingreagent, such as diphenylphosphoryl azide, in the presence of anappropriate base, such as triethylamine, at elevated temperature, in anappropriate solvent, such as toluene, to give compound 10. Compound 24is reacted with compound 10 in the presence of an appropriate base, suchas diisopropylethylamine, in an appropriate solvent, such asdichloromethane, to give compound 25. Compound 25 is treated with anappropriate ketal deprotecting reagent, such as hydrochloric acid, in anappropriate solvent, such as a combination of water and methanol, togive a compound 26 of Formula I.

Deuterium can be incorporated to different positions synthetically,according to the synthetic procedures as shown in Scheme I, by usingappropriate deuterated intermediates. For example, to introducedeuterium at one or more positions of R₁₆-R₂₁, compound 28 with thecorresponding deuterium substitutions can be used. To introducedeuterium at R₁₃-R₁₅, compound 41 with the corresponding deuteriumsubstitutions can be used. To introduce deuterium at R₁₁, deuteriumoxide and/or d₆-deuterium oxide can be used. To introduce deuterium atR₁₂, d₄-methanol can be used. To introduce deuterium at one or morepositions of R₉-R₁₀ or R₂₄-R₂₇, lithium aluminum deuteride can be used.To introduce deuterium at one or more positions of R₁-R₄ and R₇,compound 2 with the corresponding deuterium substitutions can be used.To introduce deuterium at one or more positions of R₅-R₆, compound 50with the corresponding deuterium substitutions can be used.

Deuterium can be incorporated to various positions having anexchangeable proton, such as the amine N—H and hydroxyl O—Hs, viaproton-deuterium equilibrium exchange. For example, to introducedeuterium at R₈, R₂₂-R₂₃, and R₂₈, this proton may be replaced withdeuterium selectively or non-selectively through a proton-deuteriumexchange method known in the art.

The invention is further illustrated by the following examples. AllIUPAC names were generated using CambridgeSoft's ChemDraw 10.0.

EXAMPLE 1(1S,2S,3R,5S)-3-(7-((1R,2S)-2-(3,4-difluorophenyl)cyclopropylamino)-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl)-5-(2-hydroxyethoxy)cyclopentdi-1,2-diol(ticagrelor)

Step 1

(R)-tert-Butyl 1-(hydroxyamino)-1-oxopropan-2-ylcarbamate

At about 0° C., isopropyl chloroformate (86.05 g, 702.16 mmol, 1.00equiv.) was added dropwise, over a period of 60 minutes, to a stirredmixture of (S)-2-(tert-butoxycarbonylamino)propanoic acid (118 g, 623.65mmol, 1.00 equiv.), tetrahydrofuran (500 mL), and triethylamine (63.63g, 628.82 mmol, 1.00 equiv.). The resulting mixture was then stirred atabout 0° C. for about 2 hours, and then the solids were removed byfiltration. The resulting filtrate was then added to a hydroxylaminesolution (formed by first stirring a mixture of sodium hydroxide (37.6g, 940.00 mmol, 1.50 equiv.), methanol (500 mL), and hydroxylaminehydrochloride (65 g, 935.39 mmol, 1.50 equiv) at about 0° C. for about 2hours, and then removing the resulting solids by filtration). Theresulting mixture was stirred at about 0° C. for about 2 hours, thesolids were removed by filtration, and the resulting filtrate wasconcentrated in vacuo. The resulting residue was purified by silica gelcolumn chromatography (ethyl acetate/petroleum ether (1:1)) to give thetitle product as a white solid (70 g; yield=55%). ¹H NMR (300 MHz,CDCl₃) δ: 5.18 (b, 1H), 4.21 (m, 1H), 2.06 (s, 1H), 1.46 (s, 9H), 1.40(d, J=7.2 Hz, 3H).

Step 2

tert-Butyl-(R)-1-(3-oxa-2-aza-bicyclo[2.2.1]hept-5-en-2-yl)-1-oxopropan-2-ylcarbamate

At about −78° C. and under an atmosphere of nitrogen, oxalyl dichloride(30.43 g, 239.74 mmol, 4.00 equiv.) was added dropwise, over a period of20 minutes, to stirred solution of dimethylsulfoxide (28.1 g, 359.66mmol, 6.00 equiv.) in dichloromethane (200 mL). To this mixture wasadded dropwise, over a period of about 10 minutes, a solution of(R)-tert-butyl 1-(hydroxyamino)-1-oxopropan-2-ylcarbamate (12.24 g,59.94 mmol, 1.00 equiv.) and cyclopenta-1,3-diene (4.15 g, 62.78 mmol,1.05 equiv.) in a mixture of dichloromethane/dimethylsulfoxide (5:1) (60mL). The resulting mixture was stirred for at about −78° C. for about 30minutes, and then triethylamine (66.8 mL) was added. The resultingmixture was washed with 1M hydrochloric acid (2×200 mL), and extractedwith dichloromethane (3×100 mL). After the organic layers were combined,the organic phase was washed with 10% sodium bicarbonate (2×200 mL),dried over anhydrous sodium sulfate, and concentrated in vacuo. Theresulting residue was purified by silica gel column chromatography(ethyl acetate/petroleum ether (1:10)) to give the desired product as awhite solid (8.6 g; yield=53%). ¹H NMR (300 MHz, CDCl₃) δ: 6.54 (s, 1H),6.39 (m, 1H), 5.22-5.34 (m, 3H), 4.51 (m, 1H), 2.02 (d, J=8.4 Hz, 1H),1.86 (d, J=8.7 Hz, 1H), 1.43 (s, 9H), 1.09 (m, 3H).

Step 3

tert-Butyl(R)-1-(5,6-dihydroxy-3-oxa-2-aza-bicyclo[2.2.1]heptan-2-yl)-1-oxopropan-2-ylcarbamate

A solution oftert-butyl-(R)-1-(3-oxa-2-aza-bicyclo[2.2.1]hept-5-en-2-yl)-1-oxopropan-2-ylcarbamate(27.3 g, 101.75 mmol, 1.00 equiv.) in teterahydrofuran:water (5:1) (600mL), osmium tetroxide (230 mg, 0.90 mmol, 0.01 equiv.), andN-methylmorpholine-N-oxide (25.26 g, 215.62 mmol, 2.10 equiv.) wasstirred at about 20° C. for about 50 minutes. After adding sodiumthiolsulfate (22 g), the resulting solution was extracted with ethylacetate (3×200 mL) and the organic layers combined. The organic phasewas washed with a saturated sodium bicarbonate solution (1×250 mL) andthen concentrated in vacuo to give the desired product as a yellowliquid (30.1 g; yield=98%). ¹H NMR (300 MHz, CDCl₃) δ: 5.29 (b, 1H),4.36-4.74 (m, 3H), 3.76-4.09 (m, 4H), 2.67 (m, 1H), 1.90 (m, 1H), 1.43(s, 9H), 1.30 (d, J=6.9 Hz, 3H).

(2-(4,4-Dimethyl-3,5,8-trioxa-9-aza-tricyclo[5.2.1.0^(2,b)]dec-9-yl)-1-methyl-2-oxo-ethyl]-carbamicacid tert-butyl ester

A solution of tert-butyl(R)-1-(5,6-dihydroxy-3-oxa-2-aza-bicyclo[2.2.1]heptan-2-yl)-1-oxopropan-2-ylcarbamate(30.1 g, 99.56 mmol, 1.00 equiv.) in 2,2-dimethoxypropane (600 mL) andp-toluenesulfonic acid (1.1 g, 6.39 mmol, 0.06 equiv.) was stirred atabout 22° C. for about 50 minutes. After adding a saturated solution ofsodium bicarbonate (450 mL), the mixture was extracted with ethylacetate (3×300 mL). The organic layers were combined and concentrated invacuo to give the title product as a white solid (33.5 g; yield=98%). ¹HNMR (300 MHz, CDCl₃) δ: 5.22 (b, 1H), 4.89 (s, 1H), 4.70 (s, 1H), 4.61(b, 1H), 4.33 (s, 2H), 2.24 (d, J=11.4 Hz, 1H), 1.82 (m, 1H), 1.52 (s,3H), 1.46 (s, 9H), 1.32 (d, J=6.9 Hz, 3H), 1.27 (s, 3H).

Step 5

4,4-Dimethyl-3,5,8-trioxa-9-aza-tricyclo[5.2.1.0^(2,6)]decane

A mixture of[2-(4,4-dimethyl-3,5,8-trioxa-9-aza-tricyclo[5.2.1.0^(2,b)]dec-9-yl)-1-methyl-2-oxo-ethyl]-carbamicacid tert-butyl ester (33.5 g, 97.84 mmol, 1.00 equiv.), methanol (500mL), and sodium borohydride (14.67 g, 388.10 mmol, 4.00 equiv) wasstirred at about 20° C. for about 50 minutes. The pH value of themixture was then adjusted to 3 by adding 1M hydrochloric acid. Afterextracting the mixture with ethyl acetate (200 mL), the aqueous layerswere combined and the pH was adjusted to 10 by adding a 10% sodiumbicarbonate solution. Standard extractive workup with ethyl acetate(3×300 mL) gave the title product as a white solid (16.7 g;yield=99.8%). ¹H NMR (300 MHz, CDCl₃) δ: 4.80 (b, 1H), 4.69 (s, 1H),4.29 (d, J=5.4 Hz, 1H), 4.22 (d, J=5.4 Hz, 1H), 3.77 (s, 1H), 2.28 (d,J=11.1 Hz, 1H), 1.65 (d, J=11.4 Hz, 1H), 1.50 (s, 3H), 1.27 (s, 3H).

Step 6

(3aR,4S,6R,6aS)-6-amino-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-ol

Under a presurrized hydrogen atmosphere (3 atm), a suspension of4,4-dimethyl-3,5,8-trioxa-9-aza-tricyclo[5.2.1.0^(2,6)]decane (16.7 g,97.55 mmol, 1.00 equiv.), 10% palladium on carbon (1.67 g), and methanol(250 mL) was stirred at about 20° C. for about 60 minutes. Afterfiltering the solution, the resulting filtrate was concentrated in vacuoto give the title product as a white solid (16.8 g; yield=99%). MS:m/z=174 (MH)⁺.

Step 7

Benzyl(3aS,4R,6S,6aR)-6-hydroxy-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-ylcarbamate

At about 0° C., benzyl carbonochloridate (18.4 g, 107.86 mmol, 1.05equiv.) was added to the solution of(3aR,4S,6R,6aS)-6-amino-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-ol(17.0 g, 98.15 mmol, 1.00 equiv.) and sodium carbonate (20.8 g, 196.24mmol, 2.00 equiv) in tetrahydrofuran:water (5:1) (600 mL). Standardextractive workup with ethyl acetate (3×200 mL) gave the title productas a white solid (20.5 g; yield=68%). ¹H NMR (300 MHz, CDCl₃) δ:7.30-7.39 (m, 5H), 5.20 (s, 2H), 4.60 (d, J=5.4 Hz, 1H), 4.50 (d, J=5.4Hz, 1H), 4.27 (s, 1H), 4.19 (d, J=5.7 Hz 1H), 2.26 (m, 1H), 1.71 (d,J=14.4 Hz, 1H), 1.47 (s, 3H), 1.28 (s, 3H).

Step 8

Ethyl2-((3aR,4S,6R,6aS)-6-(benzyloxycarbonyl)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)acetate

Benzyl(3aS,4R,6S,6aR)-6-hydroxy-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-ylcarbamate(12.29 g, 39.99 mmol, 1.00 equiv.) was added to a solution of sodiumhydride (70%) (1.44 g, 60.00 mmol, 1.05 equiv.) in dimethylformamide(200 mL). The solution was stirred at about −30° C. for about 30minutes, and then ethyl 2-bromoacetate (7.68 g, 45.99 mmol, 1.20 equiv)was added. The resulting solution was stirred at ambient temperature forabout 5.5 hours, and then water was added (500 mL). Following standardextractive workup with ethyl acetate (3×200 mL), the resulting residuewas purified by silica gel column chromatography (ethyl acetate:petroleum ether (1:10)) to afford the title product as a colorless solid(0.9 g; yield=69%). ¹H NMR (300 MHz, CD₃OD) δ: 7.30-7.35 (m, 5H), 5.07(s, 2H), 4.60 (d, J=5.7 Hz, 1H), 4.51 (d, J=5.7 Hz, 1H), 4.12-4.29 (q,J=16.5 Hz, 2H), 4.10-4.22 (q, J=7.2 Hz, 2H), 4.00 (d, J=5.7 Hz, 1H),3.92 (J=4.3 Hz, 1H), 2.17-2.19 (m, 1H), 1.84 (d, J=14.7 Hz, 1H), 1.38(s, 3H), 1.26 (s, 3H), 1.11-1.22 (t, J=7.2 Hz, 3H).

Step 9

Ethyl2-((3aR,4S,6R,6aS)-6-amino-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)acetate

Under an atmosphere of hydrogen, a suspension of ethyl2-((3aR,4S,6R,6aS)-6-(benzyloxycarbonyl)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)acetate(150 mg, 0.38 mmol, 30.00 equiv.) and 10% palladium on carbon (16 mg,0.15 mmol, 1.00 equiv) in methanol (10 mL) was stirred at ambienttemperature for about 80 minutes. After filtering, the resultingfiltrate was concentrated in vacuo to give the title product as a yellowsolid (80 mg; yield=82%). ¹H NMR (300 MHz, CDCl₃) δ: 4.64 (s, 1H), 4.42(s, 1H), 4.08-4.20 (m, 4H), 3.87 (s, 1H), 3.31 (s, 1H), 2.17 (m, 1H),1.76 (m, 1H), 1.36 (s, 3H), 1.21-1.29 (m, 6H).

Step 10

2-((3aR,4S,6R,6aS)-6-amino-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol

A solution of ethyl2-((3aR,4S,6R,6aS)-6-amino-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)acetate(4.2 g, 16.2 mmol, 1.00 equiv.) in dry tetrahydrofuran (50 mL) wasslowly added to a suspension of lithium aluminum hydride (1.23 g, 32.4mmol, 2.00 equiv.) in tetrahydrofuran (50 mL). The mixture was heated atreflux for about 1 hour, and then water was added (2 mL). After thesolids were collected by filtratration, the solids were washed withtetrahydrofuran (50 mL) and then dried in vacuo to give the titleproduct as a yellow oil (2.3 g, 65%). MS: m/z=218 (MH)⁺.

Step 11

2-(Propylthio)pyrimidine-4,6-diol

A solution of 2-mercaptopyrimidine-4,6-diol (25 g, 173.61 mmol, 1.00equiv.), sodium hydroxide (15.8 g, 395.00 mmol, 2.27 equiv.),1-methylpyrrolidin-3-one (50 mL), and 1-iodopropane (30.6 g, 180.00mmol, 1.05 equiv.) dissolved in water (60 mL) was stirred at ambienttemperature for about 48 hours. The pH value of the solution wasadjusted to 2-3 by adding hydrochloric acid. The solids were thencollected by filtration to give the product as an off-white solid (35 g;(crude)). ¹H NMR (300 MHz, DMSO-d₆) δ: 11.78 (b, 1H), 10.30 (b, 1H),5.13 (s, 1H), 3.07 (t, J=7.2 Hz, 2H), 1.58-1.70 (m, 2H), 0.96 (t, J=7.2Hz, 2H).

Step 12

5-Nitro-2-(propylthio)pyrimidine-4,6-diol

A solution of 2-(propylthio)pyrimidine-4,6-diol (3 g, 16.11 mmol, 1.00equiv.) in nitric acid (65%) (10 mL) was stirred at ambient temperaturefor about 2 hours. After adding water (10 mL), the mixture was stirredat about 0° C. for about 30 minutes. The resulting solids were collectedby filtration to afford the title product as a yellow solid (1.8 g;yield=48%). ¹H NMR (300 MHz, DMSO-d₆) δ: 3.17 (t, J=7.2 Hz, 2H),1.62-1.75 (m, 2H), 0.95-1.02, (t, J=7.2 Hz, 3H).

Step 13

4,6-Dichloro-5-nitro-2-(proplthio)pyrimidine

A solution of 5-nitro-2-(propylthio)pyrimidine-4,6-diol (1.8 g, 6.71mmol, 1.00 equiv.), phosphoryl chloride (15 mL) andN,N-diethylbenzenamine (2 mL) was stirred at reflux for about 3 hours inan oil bath. After cooling the mixture to about 20° C. with a water/icebath, the mixture was concentrated in vacuo. The resulting residue waspurified by silica gel column chromatography (ethyl acetate/petroleumether (1:100)) to give the title product as a yellow oil (1.1 g;yield=61%). ¹H NMR (300 MHz, CDCl₃) δ: 3.15 (t, J=7.2 Hz, 2H), 1.73-1.87(m, 2H), 1.07-1.13 (J=7.2 Hz, 3H).

Step 14

2-((3aR,4S,6R,6aS)-6-(6-Chloro-5-nitro-2-(propylthio)pyrimidin-4-ylamino)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol

A solution of 4,6-dichloro-5-nitro-2-(propylthio)pyrimidine (1.07 g,3.99 mmol, 1.00 equiv.), and2-((3aR,4S,6R,6aS)-6-amino-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol(570 mg, 4.41 mmol, 1.20 equiv.) in tetrahydrofuran (20 mL) was stirredat 0-10° C. for about 2 hours and then water (20 mL) was added. Standardextractive workup with ethyl acetate (3×20 mL) afforded the titleproduct as a yellow oil (800 mg; yield=45%). ¹H NMR (300 MHz, CDCl₃) δ:8.65 (b, 1H), 4.66-4.76 (m, 2H), 4.56 (m, 1H), 3.99 (d, J=7.5 Hz, 1H),3.70-3.87 (m, 3H), 3.64-3.67 (m, 1H), 3.07-3.20 (m, 2H), 2.34 (m, 1H),1.97 (m, 1H), 1.76-1.82 (m, 2H), 1.46 (s, 3H), 1.27 (s, 3H), 1.07 (t,J=7.5 Hz, 3H).

Step 15

2-((3aR,4S,6R,6aS)-6-(5-Amino-6-chloro-2-(propylthio)pyrimidin-4-ylamino)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol

A suspension of2-((3aR,4S,6R,6aS)-6-(6-chloro-5-nitro-2-(propylthio)pyrimidin-4-ylamino)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol(800 mg, 1.78 mmol, 1.00 equiv.), iron powder (800 mg, 14.29 mmol, 8.00equiv.), acetic acid (860 mg, 14.33 mmol, 8.00 equiv.) and water/ethanol(10 mL) was stirred at about 60° C. for about 20 minutes in an oil bath.After the solids were removed by filtration, the resulting filtrate wasextracted with dichloromethane (3×10 mL). The organic layers werecombined, dried over anhydrous sodium sulfate, and concentrated in vacuoto give the title product as a yellow oil (780 mg; yield=93%). MS:m/z=419 (MH)⁺.

Step 16

2-((3aR,4S,6R,6aS)-6-(7-Chloro-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol

A solution of sodium nitrite (148 mg, 2.14 mmol, 1.12 equiv.) in water(1 mL) was added to a solution of2-((3aR,4S,6R,6aS)-6-(5-amino-6-chloro-2-(propylthio)pyrimidin-4-ylamino)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol(800 mg, 1.91 mmol, 1.00 equiv.) and acetic acid (680 mg, 11.33 mmol,5.90 equiv.) in toluene (9 mL). The resulting solution was stirred atabout 20° C. for about 30 minutes, and then the pH value of the solutionwas adjusted to 8-9 by adding potassium carbonate. Following standardextractive workup with ethyl acetate (3×10 mL), the resulting residuewas purified by silica gel column (ethyl acetate/petroleum ether (1:10))to give the title product as a yellow oil (370 mg; yield=45%). ¹H NMR(300 MHz, CDCl₃) δ: 5.54-5.56 (q, J₁=2.4 Hz, J2=6.3 Hz, 1H), 5.21-5.25(m, 1H), 4.90 (d, J=6.3 Hz, 1H), 4.05-4.09 (m, 1H), 3.50-3.66 (m, 4H),3.23 (t, J=7.5 Hz, 2H), 2.68-2.72 (m, 1H), 2.58 (m, 1H), 1.81-1.89 (m,2H), 1.57 (s, 3H), 1.39 (s, 3H), 1.12, (t, J=7.5 Hz, 3H).

Step 17

3-(3,4-Difluoro-phenyl)-acryloyl chloride

At 0-5° C., oxalyl dichloride (25.7 g, 202.36 mmol, 3.00 equiv.) wasadded to a mixture of (E)-3-(3,4-difluorophenyl)acrylic acid (12.4 g,67.38 mmol, 1.00 equiv.), N,N-dimethylformamide (1 mL), anddichloromethane (150 mL). The solution was stirred at ambienttemperature for about 2 hours, and then concentrated in vacuo to givethe title product, which was used in the next step without any furtherpurification.

Step 17

[3aS-[1(E),3a,6,7-a]]-1-[3-(3,4-Difluorophenyl)-1-oxo-2-propenyl]-hexahydro-8,8-dimethyl-3H-3a,6-methano-2,1-benzisothiazole-2,2-dioxide

At 0-5° C., a solution of 3-(3,4-difluoro-phenyl)-acryloyl chloride indichloromethane (30 mL) was added to a mixture of(2R)-bornane-10,2-sultam (14.5 g, 67.35 mmol, 1.00 equiv), triethylamine(20.4 g, 201.98 mmol, 3.00 equiv), and dichloromethane (120 mL). Theresulting solution was stirred at ambient temperature for about 3 hoursand then water (40 mL) was added. Standard extractive workup withdichloromethane (2×40 mL) gave the title product as an off-white solid(18.5 g; yield=72%). ¹H NMR (300 MHz, CDCl₃) δ: 7.68 (d, J=15.6 Hz, 1H),7.15-7.45 (m, 3H), 7.19 (d, J=15.6 Hz, 1H), 4.00 (m, 1H), 3.55 (q,J₁=13.8 Hz, J₂=24.0 Hz), 2.18 (m, 2H), 1.93 (m, 2H), 1.37-1.48 (m, 2H),1.22 (s, 3H), 0.96 (s, 3H).

Step 18

[3aS-[1(1R,2R),3a,6,7-a]]-1-[[2-(3,4-Difluorophenyl)cyclopropyl]carbonyl]-hexahydro-8,8-dimethyl-3H-3a,6-methano-2,1-benzisothiazole-2,2-dioxide

At 0-5° C., 1-methyl-1-nitrosourea (39.1 g, 379.61 mmol, 2.50 equiv.)was added in portions to a mixture of 50% aqueous sodium hydroxide (150mL) and ethyl ether (300 mL). After the solid was dissolved, the aqueousphase was removed. At 0-5° C., a solution of[3aS-[1(E),3a,6,7a]]-1-[3-(3,4-difluorophenyl)-1-oxo-2-propenyl]-hexahydro-8,8-dimethyl-3H-3a,6-methano-2,1-benzisothiazole-2,2-dioxide(48.2 g, 126.51 mmol, 1.00 equiv.), palladium(II) acetate (200 mg, 1.04mmol) in dichloromethane (300 mL) was then added to the mixture. Themixture was stirred at ambient temperature for about 1 hour, acetic acid(100 mL) was added, and then water (500 mL) was added. Standardextractive workup with dichloromethane (2×100 mL) afforded the titleproduct as a light-yellow oil (48 g; yield=80%). ¹H NMR (300 MHz, CDCl₃)δ: 6.97-7.09 (m, 3H), 3.91-3.95 (m, 1H), 3.44-3.57 (q, J₁=13.8 Hz,J₂=24.9 Hz), 2.56 (m, 2H), 2.15 (m, 2H), 1.90-1.95 (m, 3H), 1.76-1.82(m, 1H), 1.22-1.47 (m, 3H), 1.20 (s, 3H), 1.00 (s, 3H).

Step 19

(1R,2R)-2-(3,4-Difluorophenyl)cyclopropanecarboxylic acid

A mixture of[3aS-[1(1R,2R),3a,6,7a]]-1-[[2-(3,4-difluorophenyl)cyclopropyl]carbonyl]-hexahydro-8,8-dimethyl-3H-3a,6-methano-2,1-benzisothiazole-2,2-dioxide(48 g, 121.52 mmol, 1.00 equiv.) in 10% lithium hydroxide (200 mL) andtetrahydrofuran (200 mL) was stirred at about 50° C. for about 0.5hours. The mixture was cooled to ambient temperature, and washed ether(2×100 mL). The pH value of the aqueous layer was adjusted to 3 byadding 12N hydrochloric acid. Following standard extractive workup withether (3×200 mL), the crude residue was purified by silica gel columnchromatography (ethyl acetate/petroleum ether (1:4)) to give the titleproduct as a white solid (16.0 g; yield=66%). ¹H NMR (300 MHz, CDCl₃) δ:7.05-7.14 (m, 1H), 6.85-6.96 (m, 2H), 2.54-2.60 (m, 1H), 1.84-1.90 (m,1H), 1.65-1.72 (m, 1H), 1.30-1.40 (m, 1H).

Step 20

(1R,2S)-2-(3,4-Difluorophenyl)cyclopropanamine

A solution of (1R,2R)-2-(3,4-difluorophenyl)cyclopropanecarboxylic acid(8.0 g, 40.40 mmol, 1.00 equiv.), diphenylphosphoryl azide (11.2 g,40.73 mmol, 1.00 equiv.), triethylamine (6.2 g, 61.39 mmol, 1.50 equiv.)in toluene (60 mL) was heated at reflux for about 1 hour and thenrefluxing 6N hydrogen chloride was added. The mixture was kept at refluxfor 16 hours, and then cooled to ambient temperature. The resultingmixture was concentrated in vacuo, and the resulting residue wasdissolved in water/ether (1:1) 200 mL). Following standard extractiveworkup with ether (3×100 mL), the resulting residue was purified bysilica gel column chromatography (ethyl acetate/petroleum ether(1:4˜1:0)) to give the title product as a light-brown solid (6.2 g;yield=91%). ¹H NMR (300 MHz, CDCl₃) δ: 6.98-7.07 (m, 1H), 6.72-6.81 (m,2H), 2.40 (m, 1H), 1.82-1.87 (m, 1H), 1.05-1.11 (m, 1H), 0.85-0.96 (m,1H).

Step 21

2-((3aR,4S,6R,6aS)-6-(7-((1R,2S)-2-(3,4-Difluorophenyl)cyclopropylamino)-5-(propylthio)-3H-[1,2,3-]triazolo[4,5-d]pyrimidin-3-yl)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol

A solution of2-((3aR,4S,6R,6aS)-6-(7-chloro-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol(370 mg, 0.86 mmol, 1.00 equiv) (15 mL),(1R,2S)-2-(3,4-difluorophenyl)cyclopropanamine (145.6 mg, 0.86 mmol,1.00 equiv.) and N,N-diisopropylethylamine (155.7 mg, 1.21 mmol, 1.20equiv.) in dichloromethane was stirred at ambient temperature for about16 hours, and then water (10 mL) was added. Standard extractive workupwith dichloromethane (3×10 mL) gave the title product as a yellow oil(450 mg; yield=93%). m/z=563 (MH)⁺.

Step 22

(1S,2S,3R,5S)-3-(7-((1R,2S)-2-(3,4-difluorophenyl)cyclopropylamino)-5-(propylthio)-3H-[1,2,3-]triazolo[4,5-d]pyrimidin-3-yl)-5-(2-hydroxyethoxy)cyclopentane-1,2-diol(ticagrelor-d₀)

A solution of2-((3aR,4S,6R,6aS)-6-(7-((1R,2S)-2-(3,4-difluorophenyl)cyclopropylamino)-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol(450 mg, 0.80 mmol, 1.00 equiv.) in methanol (4 mL) and 12N hydrochloricacid (1.5 mL) was stirred at ambient temperature for about 3 hours. ThepH value of the solution was adjusted to 8-9 by adding potassiumcarbonate. Following standard extractive workup with ethyl acetate (3×20mL), the resulting crude residue was purified by silica gel columnchromatography (dichloromethane/methanol (50:1)) to give a semi-crudeproduct (200 mg; yield=48%). The semi-crude product, which containedabout 5% of other diastereoisomers, was then further purified bychiral-prep HPLC (column: Chiralpak IA2×25 cm, 5umChiral-P(IA)004IA00CJ-MB003) to afford the title compound (100 mg).[α]_(D) ^(24.1) −43.2° (c, 0.2 g/100 mL in MeOH). LC-MS: m/z=523.0(MH)⁺, Retention time: 1.58 minute. ¹H NMR (300 MHz, CD₃OD) δ: 7.08-7.23(m, 3H), 5.13 (q, 1H), 4.75-4.79 (m, 1H), 4.17-4.20 (m, 1H), 3.91-3.95(m, 1H), 3.63-3.73 (m, 4H), 3.06-3.26 (m, 2H), 2.90-3.00 (m, 1H),2.70-2.80 (m, 1H), 2.05-2.29 (m, 2H), 1.60-1.88 (m, 2H), 1.38-1.59 (m,2H), 0.94 (t, J=7.5 Hz, 3H).

EXAMPLE 2(1S,2S,3R,5S)-3-(7-((1R,2S)-2-(3,4-difluorophenyl)cyclopropylamino)-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl)-5-(2-hydroxy-d₄-ethoxy)cyclopentane-1,2-diol(ticagrelor-d₄)

Step 1

d₄-Ethane-1,2-diol

A solution of diethyloxalate (6.5 g, 44.48 mmol, 1.00 equiv.) in drytetrahydrofuran (100 mL) was slowly added to a slurry of lithiumaluminum deuteride (1.87 g, 44.48 mmol, 1.00 equiv) in tetrahydrofuran(100 mL). The mixture was heated at reflux for about 3 hours, and thenwater (4 mL) was added. The solids were removed by filtration, and theresulting filtrate was then washed with tetrahydrofuran (100 mL). Thesolvent was removed in vacuo to give the title product as a colorlessoil (2.1 g; yield=71%).

Step 2

2-(Benzyloxy)-d₄-ethanol

Silver oxide (11.05 g, 47.72 mmol, 1.50 equiv.), and benzylbromide (5.98g, 34.99 mmol, 1.10 equiv.) were added to a stirred solution ofd₄-ethane-1,2-diol (2.1 g, 31.81 mmol, 1.00 equiv.) in dichloromethane(40 mL). The mixture was stirred at ambient temperature for about 16hours, and then filtered through a small pad of silica gel. Followingstandard extractive workup with ethyl acetate, the crude residue waspurified by silica gel column chromatography (ethyl acetate: petroleumether (1:10)) to give the title product as a colorless oil (2.85 g;yield=57%). ¹H NMR (300 MHz, CDCl₃) δ: 7.29-7.42 (m, 5H), 4.59 (s, 2H).

Step 3

1-((2-Bromo-d₄-ethoxy)methyl)benzene

At about −20° C., triphenylphosphine (5.73 g, 21.87 mmol, 1.20 equiv.)was added in portions, over a period of 15 minutes, to a solution ofd₄-2-(benzyloxy)ethanol (2.85 g, 18.24 mmol, 1.00 equiv), andN-bromosuccinimide (4.85 g, 27.25 mmol, 1.50 equiv.) in tetrahydrofuran(80 mL). The resulting solution was stirred at 15-25° C. for about 30minutes. Following standard extractive workup with ethyl acetate, theresulting crude residue was purified by silica gel column chromatography(ethyl acetate/petroleum ether (1:5)) to give the title product as acolorless liquid (2.64 g; yield=66%). ¹H NMR (300 MHz, CDCl₃) δ:7.31-7.40 (m, 5H), 4.62 (s, 2H)

Step 4

Benzyl(3aS,4R,6S,6aR)-6-(2-(benzyloxy)-d₄-ethoxy)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-ylcarbamate

At about −10° C., benzyl(3aS,4R,6S,6aR)-6-hydroxy-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-ylcarbamate(3.25 g, 10.57 mmol, 1.00 equiv.) was added to a solution of 70% sodiumhydride (0.38 g, 11.11 mmol, 1.05 equiv.) in N,N-dimethylformamide (50mL). The solution was stirred at about −10° C. for about 30 minutes, andthen 1-((2-bromo-d₄-ethoxy)methyl)benzene (2.64 g, 12.04 mmol, 1.14equiv.) was added. The resulting solution was stirred at ambienttemperature for about 24 hours, and then water (50 mL) was added.Following standard extractive workup with ethyl acetate (3×50 mL), theresulting crude product was purified by silica gel column chromatography(ethyl acetate:petroleum ether (1:10)) to give title product as acolorless solid (2.25 g; yield=48%).

Step 5

2-((3aR,4S,6R,6aS)-6-Amino-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)-d₄-ethanol

Under an atmosphere of hydrogen, a suspension of benzyl(3aS,4R,6S,6aR)-6-(2-(benzyloxy)-d₄-ethoxy)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-ylcarbamate(2.25 g, 5 mmol, 3.30 equiv.), 10% palladium on carbon (1.6 g, 1.5 mmol,1.00 equiv.), and methanol (50 mL) was stirred at ambient temperaturefor about 10 hours. The suspension was filtered, and the resultingfiltrate was concentrated in vacuo to give the title product as a yellowsolid (0.95 g; yield=86%). MS: m/z=222 (MH)⁺.

Step 6

2-((3aR,4S,6R,6aS)-6-(6-Chloro-5-nitro-2-(propylthio)pyrimidin-4-ylamino)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)-d₄-ethanol

The procedure of Example 1, Step 14 was followed, but substituting2-((3aR,4S,6R,6aS)-6-amino-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)-d₄-ethanol for2-((3aR,4S,6R,6aS)-6-amino-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol.The title product was isolated as a yellow oil (680 mg; yield=47.39%).¹H NMR (300 MHz, CDCl₃) δ: 8.66 (b, 1H), 4.65-4.76 (m, 2H), 4.56 (m,1H), 3.99 (d, J=7.5 Hz, 1H), 3.07-3.21 (m, 2H), 2.34 (m, 1H), 1.98 (m,1H), 1.77-1.82 (m, 2H), 1.46 (s, 3H), 1.27 (s, 3H), 1.06 (t, J=7.5 Hz,3H).

Step 7

2-((3aR,4S,6R,6aS)-6-(5-Amino-6-chloro-2-(propylthio)pyrimidin-4-ylamino)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)-d₄-ethanol

The procedure of Example 1, Step 15 was followed, but substituting2-((3aR,4S,6R,6aS)-6-(6-chloro-5-nitro-2-(propylthio)pyrimidin-4-ylamino)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)-d₄-ethanolfor2-((3aR,4S,6R,6aS)-6-(6-chloro-5-nitro-2-(propylthio)pyrimidin-4-ylamino)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol.The title product was isolated as a yellow oil (700 mg (crude)). MS:m/z=423 (MH)⁺.

Step 8

2-((3aR,4S,6R,6aS)-6-(7-Chloro-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)-d₄-ethanol

The procedure of Example 1, Step 16 was followed but substituting2-((3aR,4S,6R,6aS)-6-(5-amino-6-chloro-2-(propylthio)pyrimidin-4-ylamino)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)-d₄-ethanolfor2-((3aR,4S,6R,6aS)-6-(5-amino-6-chloro-2-(propylthio)pyrimidin-4-ylamino)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol.The title product was isolated as a yellow oil (800 mg; (crude)). ¹H NMR(300 MHz, CDCl₃) δ: 5.53-5.57 (q, J₁=2.4 Hz, J2=6.3 Hz, 1H), 5.22-5.25(m, 1H), 4.91 (d, J=6.3 Hz, 1H), 4.05-4.09 (m, 1H), 3.25 (t, J=7.5 Hz,2H), 2.68-2.72 (m, 1H), 2.57 (m, 1H), 1.81-1.88 (m, 2H), 1.57 (s, 3H),1.39 (s, 3H), 1.11, (t, J=7.5 Hz, 3H).

Step 9

2-((3aR,4S,6R,6aS)-6-(7-((1R,2S)-2-(3,4-Difluorophenyl)cyclopropylamino)-5-(propylthio)-3H-[1,2,3-]triazolo[4,5-d]pyrimidin-3-yl)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)-d₄-ethanol

The procedure of Example 1, Step 21 was followed but substituting2-((3aR,4S,6R,6aS)-6-(7-chloro-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)-d₄-ethanolfor2-((3aR,4S,6R,6aS)-6-(7-chloro-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol.The title product was isolated as a yellow oil (450 mg; yield=43%). MS:m/z=567 (MH)⁺.

Step 10

(1S,2S,3R,5S)-3-(7-((1R,2S)-2-(3,4-Difluorophenyl)cyclopropylamino)-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl)-5-(2-hydroxy-d₄-(ethoxy)cyclopentane-1,2-diol(ticagrelor-d₄)

The procedure of Example 1, Step 22 was followed but substituting2-((3aR,4S,6R,6aS)-6-(7-((1R,2S)-2-(3,4-difluorophenyl)cyclopropylamino)-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)-d₄-ethanolfor2-((3aR,4S,6R,6aS)-6-(7-((1R,2S)-2-(3,4-difluorophenyl)cyclopropylamino)-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol.The title product was first isolated as an semi-pure off-white solid(300 mg) that was then further purified by chiral-prep HPLC (column:Chiralpak IA2×25 cm, 5 umChiral-P(IA)004IA00CJ-MB003) to give the titlecompound as a nearly pure product (210 mg; yield=50%). [α]_(D) ^(24.1)−19.0° (c, 0.1 g/100 mL in MeOH). LC-MS: m/z=527.0 (MH)⁺, Retentiontime: 1.58 minute. ¹H NMR (300 MHz, CD₃OD) δ: 7.09-7.24 (m, 3H), 5.14(q, 1H), 4.75-4.80 (m, 1H), 4.17-4.20 (m, 1H), 3.90-3.95 (m, 1H),3.06-3.26 (m, 2H), 2.90-3.00 (m, 1H), 2.70-2.81 (m, 1H), 2.05-2.30 (m,2H), 1.61-1.88 (m, 2H), 1.38-1.59 (m, 2H), 0.94 (t, J=7.5 Hz, 3H).

EXAMPLE 3(1S,2S,3R,5S)-3-(7-((1R,2S)-2-(3,4-Difluorophenyl)cyclopropylamino)-7-propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl)-5-(2-hydroxyethoxy)cyclopentane-1,2-diol(ticagrelor-d₇)

Step 1

d₃-Diethyl 2-methylmalonate

Sodium metal (7.59 g, 330.00 mmol, 1.05 equiv.) was slowly added toethanol (500 mL) and stirred at ambient temperature until all the sodiummetal was consumed. At about 0° C., diethyl malonate (50 g, 312.50 mmol,1.00 equiv.) was added dropwise, over a period of 30 minutes, to thestirred solution. At about 0° C., iodomethane-d₃ (47.85 g, 330.00 mmol,1.05 equiv) was then added dropwise, over a period of about 2 hours, tothe stirred solution. The solution was stirred at ambient temperaturefor about 3 hours and then concentrated in vacuo. After adding water(500 mL), standard extractive workup with ethyl acetate (3×300 mL)afforded the title product as a light yellow liquid (48 g; yield=87%).¹H NMR (300 MHz, CDCl₃) δ: 4.15 (q, J=7.2 Hz, 2H), 2.30 (s, 1H), 1.28(t, J=7.2 Hz, 3H).

Step 2

d₄-Diethyl 2-methylmalonate

A solution of d₃-diethyl 2-methylmalonate (48 g, 271.19 mmol, 1.00equiv.) and triethylamine (27.4 g, 271.29 mmol, 1.00 equiv.) ind₄-methanol (240 mL) was stirred at about 25° C. for about 16 hours. Theresulting mixture was then concentrated in vacuo to give the titleproduct as a light yellow liquid (48 g; yield=99%). ¹H NMR (300 MHz,CDCl₃) δ: 4.16 (q, J=7.2 Hz, 2H), 1.29 (t, J=7.2 Hz, 3H).

Step 3

d₅-Ethyl Propionate

A mixture of d₄-diethyl 2-methylmalonate (42 g, 235.69 mmol, 1.00equiv.), sodium chloride (27.5 g, 470.57 mmol, 2.00 equiv.), deuteriumoxide (4.8 g, 240.00 mmol, 1.00 equiv.) and dimethylsulfoxide-d₆ (200mL) was stirred at 150-160° C. for about 3 hours. The solvent was thenremoved by distillation. Standard extractive workup with ethyl ether(250 mL) gave the title product, which was used in the next step withoutfurther purification (15 g; yield=59%), ¹H NMR (300 MHz, CDCl₃) δ: 4.13(q, J=7.2 Hz, 2H), 1.25 (t, J=7.2 Hz, 3H).

Step 4

d₅-Propan-1-ol

At 0-5° C., lithium aluminum deuteride (5.8 g, 138.10 mmol, 0.80 equiv.)was added to a solution of ds-ethyl propionate (18.6 g, 173.83 mmol,1.00 equiv.) in ethyl ether (200 mL). The resulting solution was stirredat ambient temperature for about 3 hours, and then deuterium oxide (50mL) was added. The solution was stirred at ambient temperature for about1 hour, and then the pH value of the solution was adjusted to 4-5 byadding 10% sulfuric acid. The crude product was purified bydistillation. The fraction collected was at 70-88° C. to give the titleproduct (19.8 g; (crude, contained water and ethanol)) as a colorlessliquid, which was used in the next step without more purification.

Step 5

d₇-1-Iodopropane

A solution of d₅-propan-1-ol (19.8 g, 295.52 mmol, 1.00 equiv.) in 45%hydroiodic acid (180 mL) was heated at reflux for about 17 hours. Theorganic phase was separated and washed with sodium sulfate (1×10 mL) andbrine (1×10 mL). The resulting crude product was then purified bydistillation (1 atm). The fraction collected was at 95-101° C. to givethe title product as a colorless liquid (5.1 g; yield=26%).

Step 6

2-(d₇-Propylthio)pyrimidine-4,6-diol

The procedure of Example 1, Step 11, was followed but substitutingd₇-1-iodopropane for 1-iodopropane. The title product was isolated as anoff-white solid (3.5 g; yield=64%).

Step 7

5-Nitro-2-(d₇-propylthio)pyrimidine-4,6-diol

The procedure of Example 1, Step 12 was followed, but substituting2-(propyl-d₇-thio)pyrimidine-4,6-diol for2-(propylthio)pyrimidine-4,6-diol. The title product was isolated as ayellow solid (3.2 g; yield=51%).

Step 8

4,6-Dichloro-5-nitro-2-(d₇-propylthio)pyrimidine

The procedure of Example 1, Step 13 was followed, but substituting5-nitro-2-(d₇-propylthio)pyrimidine-4,6-diol for5-nitro-2-(propylthio)pyrimidine-4,6-diol. The title product wasisolated as as a yellow oil (1.8 g; yield=49%).

Step 9

2-((3aR,4S,6R,6aS)-6-(6-Chloro-5-nitro-2-(d₇-propylthio)pyrimidin-4-ylamino)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]1-dioxol-4-yloxy)ethanol

The procedure of Example 1, Step 14 was followed, but substituting4,6-dichloro-5-nitro-2-(d₇-propylthio)pyrimidine for4,6-dichloro-5-nitro-2-(propylthio)pyrimidine. The title product wasisolated as a yellow oil (800 mg; yield=57%). ¹H NMR (300 MHz, CDCl₃) δ:8.66 (b, 1H), 4.66-4.77 (m, 2H), 4.56 (m, 1H), 3.99 (d, J=7.5 Hz, 1H),3.71-3.87 (m, 3H), 3.64-3.66 (m, 1H), 2.33 (m, 1H), 1.97 (m, 1H), 1.45(s, 3H), 1.26 (s, 3H).

Step 10

2-((3aR,4S,6R,6aS)-6-(5-Amino-6-chloro-2-(d₇-propylthio)pyrimidin-4-ylamino)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol

The procedure of Example 1, Step 15 was followed, but substituting2-((3aR,4S,6R,6aS)-6-(6-chloro-5-nitro-2-(d₇-propylthio)pyrimidin-4-ylamino)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol for2-((3aR,4S,6R,6aS)-6-(6-chloro-5-nitro-2-(propylthio)pyrimidin-4-ylamino)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol.The title product was isolated as a yellow oil (700 g; yield=93%).

Step 11

2-((3aR,4S,6R,6aS)-6-(7-Chloro-5-(d₇-propylthio)-3H-[1,2,3-]triazolo[4,5-d]pyrimidin-3-yl)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol

The procedure of Example 1, Step 16 was followed, but substituting2-((3aR,4S,6R,6aS)-6-(5-amino-6-chloro-2-(d₇-propylthio)pyrimidin-4-ylamino)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol for2-((3aR,4S,6R,6aS)-6-(5-amino-6-chloro-2-(propylthio)pyrimidin-4-ylamino)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol.The title product was isolated as a yellow oil (320 mg; yield=45%). ¹HNMR (300 MHz, CDCl₃) δ: 5.54-5.57 (q, J₁=2.4 Hz, J₂=6.3 Hz, 1H),5.21-5.25 (m, 1H), 4.91 (d, J=6.3 Hz, 1H), 4.05-4.08 (m, 1H), 3.50-3.65(m, 4H), 2.68-2.72 (m, 1H), 2.58 (m, 1H), 1.57 (s, 3H), 1.38 (s, 3H).

Step 12

2-((3aR,4S,6R,6aS)-6-(7-((1R,2S)-2-(3,4-Difluorophenyl)cyclopropylamino)-5-(d₇-propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol

The procedure of Example 1, Step 21 was followed, but substituting2-((3aR,4S,6R,6aS)-6-(7-chloro-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanolfor2-((3aR,4S,6R,6aS)-6-(7-chloro-5-(d₇-propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol.The title product was isolated as a yellow oil (600 mg; yield=58%).

Step 13

(1S,2S,3R,5S)-3-(7-((1R,2S)-2-(3,4-Difluorophenyl)cyclopropylamino)-5-(d₇-propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl)-5-(2-hydroxyethoxy)cyclopentane-1,2-diol(ticagrelor-d₇)

The procedure of Example 1, Step 22 was followed, but substituting2-((3aR,4S,6R,6aS)-6-(7-((1R,2S)-2-(3,4-difluorophenyl)cyclopropylamino)-5-(d₇-propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanolfor2-((3aR,4S,6R,6aS)-6-(7-((1R,2S)-2-(3,4-difluorophenyl)cyclopropylamino)-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol.The title product was isolated as a semi pure off-white solid (350 mg)that was further purified by chiral-prep HPLC (column: Chiralpak IA2×25cm, 5 umChiral-P(IA)004IA00CJ-MB003) to give the nearly pure product(220 mg; yield=40%). [α]_(D) ^(26.3) −26.8° (c, 0.31 g/100 mL in MeOH).LC-MS: m/z=530.0 (MH)⁺, Retention time: 1.58 minute. ¹H NMR (300 MHz,CD₃OD) δ: 7.08-7.23 (m, 3H), 5.13 (q, 1H), 4.75-4.79 (m, 1H), 4.17-4.20(m, 1H), 3.91-3.95 (m, 1H), 3.63-3.73 (m, 4H), 3.14 (m, 1H), 2.70-2.80(m, 1H), 2.15-2.29 (m, 2H), 1.38-1.50 (m, 2H).

EXAMPLE 4(1S,2S,3R,5S)-3-(7-((1R,2S)-2-(3,4-difluorophenyl)cyclopropylamino)-5-(d₇-propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl)-5-(2-hydroxy-d₄-ethoxy)cyclopentane-1,2-diol(ticagrelor-d₁₁)

Step 1

2-((3aR,4S,6R,6aS)-6-(6-Chloro-5-nitro-2-(d₇-propylthio)pyrimidin-4-ylamino)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)-d₄-ethanol

The procedure of Example 3, Step 9 was followed, but substituting2-((3aR,4S,6R,6aS)-6-amino-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)-d₄-ethanol for2-((3aR,4S,6R,6aS)-6-amino-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol.The title product was isolated as a yellow oil (800 mg; yield=57%). ¹HNMR (300 MHz, CDCl₃) δ: 8.65 (b, 1H), 4.66-4.77 (m, 2H), 4.56 (m, 1H),3.99 (d, J=7.5 Hz, 1H), 2.34 (m, 1H), 1.98 (m, 1H), 1.45 (s, 3H), 1.27(s, 3H).

Step 2

d₁₁-2-((3aR,4S,6R,6aS)-6-(5-Amino-6-chloro-2-(d₇-propylthio)pyrimidin-4-ylamino)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)-d₄-ethanol

The procedure of Example 3, Step 10 was followed, but substituting2-((3aR,4S,6R,6aS)-6-(6-chloro-5-nitro-2-(d₇-propylthio)pyrimidin-4-ylamino)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)-d₄-ethanol for2-((3aR,4S,6R,6aS)-6-(6-chloro-5-nitro-2-(d₇-propylthio)pyrimidin-4-ylamino)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol.The title product was isolated as a yellow oil (560 mg; yield=81%).

Step 3

2-((3aR,4S,6R,6aS)-6-(7-chloro-5-(propyl-d₇-thio)-3H-[1,2,3-]triazolo[4,5-d]pyrimidin-3-yl)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)-d₄-ethanol

The procedure of Example 3, Step 11 was followed, but substituting2-((3aR,4S,6R,6aS)-6-(5-amino-6-chloro-2-(d₇-propylthio)pyrimidin-4-ylamino)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)-d₄-ethanolfor2-((3aR,4S,6R,6aS)-6-(5-amino-6-chloro-2-(d₇-propylthio)pyrimidin-4-ylamino)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol. The title product wasisolated as a yellow oil (460 mg; yield=80%). ¹H NMR (300 MHz, CDCl₃) δ:5.54-5.58 (q, J₁=2.4 Hz, J₂=6.3 Hz, 1H), 5.21-5.25 (m, 1H), 4.92 (d,J=6.3 Hz, 1H), 4.06-4.09 (m, 1H), 2.67-2.73 (m, 1H), 2.58 (m, 1H), 1.57(s, 3H), 1.38 (s, 3H).

Step 4

2-((3aR,4S,6R,6aS)-6-(7-((1R,2S)-2-(3,4-difluorophenyl)cyclopropylamino)-5-(d₇-propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)-d₄-ethanol

The procedure of Example 3, Step 12 was followed, but substituting2-((3aR,4S,6R,6aS)-6-(7-chloro-5-(propyl-d₇-thio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)-d₄-ethanolfor2-((3aR,4S,6R,6aS)-6-(7-chloro-5-(propyl-d-thio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol.The title product was isolated as a yellow oil (400 mg; yield=67%).

Step 5

(1S,2S,3R,5S)-3-(7-((1R,2S)-2-(3,4-difluorophenyl)cyclopropylamino)-5-(propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl)-5-(2-hydroxyethoxy)cyclopentane-1,2-diol(ticagrelor-d₁₁)

The procedure of Example 3, Step 13 was followed, but substituting2-((3aR,4S,6R,6aS)-6-(7-((1R,2S)-2-(3,4-difluorophenyl)cyclopropylamino)-5-(d₇-propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)-d₄-ethanolfor2-((3aR,4S,6R,6aS)-6-(7-((1R,2S)-2-(3,4-difluorophenyl)cyclopropylamino)-5-(d₇-propylthio)-3H-[1,2,3]triazolo[4,5-d]pyrimidin-3-yl)-2,2-dimethyl-tetrahydro-3aH-cyclopenta[d][1,3]dioxol-4-yloxy)ethanol.The title product was isolated as an semi-pure off-white solid (350 mg)that was further purified by chiral-prep-HPLC (column: Chiralpak IA2×25cm, 5 umChiral-P(IA)004IA00CJ-MB003) to give pure product 260 mg (70%).[α]_(D) ^(26.1) −23.2° (c, 0.21 g/100 mL in MeOH). LC-MS: m/z=534.0(MH)⁺, Retention time: 1.58 minute. ¹H NMR (300 MHz, CD₃OD) δ: 7.09-7.26(m, 3H), 5.14 (q, 1H), 4.75-4.79 (m, 1H), 4.17-4.20 (m, 1H), 3.91-3.95(m, 1H), 3.14 (m, 1H), 2.76-2.84 (m, 1H), 2.15-2.29 (m, 2H), 1.43-1.51(m, 2H).

The following compounds can generally be made using the methodsdescribed above. It is expected that these compounds when made will haveactivity similar to those described in the examples above.

Changes in the metabolic properties of the compounds disclosed herein ascompared to their non-isotopically enriched analogs can be shown usingthe following assays. Compounds listed above which have not yet beenmade and/or tested are predicted to have changed metabolic properties asshown by one or more of these assays as well.

Biological Activity Assays In Vitro Liver Microsomal Stability Assay

Liver microsomal stability assays were conducted at 1 mg per mL livermicrosome protein with an NADPH-generating system in 2% sodiumbicarbonate (2.2 mM NADPH, 25.6 mM glucose 6-phosphate, 6 units per mLglucose 6-phosphate dehydrogenase and 3.3 mM magnesium chloride). Testcompounds were prepared as solutions in 20% acetonitrile-water (20 μMstock solutions) and added to the assay mixture (final assayconcentration 1 μM). Final concentration of acetonitrile in the assayshould be <1%. The reactions were incubated at 37° C. Aliquots (50 μL)were taken out at times 0, 15, 30, 45, and 60 minutes, and diluted withice cold acetonitrile (200 μL) to stop the reactions. Samples arecentrifuged at 12,000 RPM for 10 minutes to precipitate proteins.Supernatants are transferred to microcentrifuge tubes and stored forLC/MS/MS analysis of the degradation half-life of the test compounds. Ithas thus been found that certain isotopically-enriched compoundsdisclosed herein that have been tested in this assay showed an increaseddegradation half-life as compared to the non-isotopically enriched drug.In certain embodiments, the increase in degradation half-life is atleast 5%; at least 10%; at least 15%; at least 20%; at least 25%; or atleast 30%.

In Vitro Metabolism Using Human Cytochrome P₄₅₀ Enzymes

The cytochrome P₄₅₀ enzymes are expressed from the corresponding humancDNA using a baculovirus expression system (BD Biosciences, San Jose,Calif.). A 0.25 milliliter reaction mixture containing 0.8 milligramsper milliliter protein, 1.3 millimolar NADP⁺, 3.3 millimolarglucose-6-phosphate, 0.4 U/mL glucose-6-phosphate dehydrogenase, 3.3millimolar magnesium chloride and 0.2 millimolar of a compound asdisclosed herein, the corresponding non-isotopically enriched compoundor standard or control in 100 millimolar potassium phosphate (pH 7.4) isincubated at 37° C. for 20 minutes. After incubation, the reaction isstopped by the addition of an appropriate solvent (e.g., acetonitrile,20% trichloroacetic acid, 94% acetonitrile/6% glacial acetic acid, 70%perchloric acid, 94% acetonitrile/6% glacial acetic acid) andcentrifuged (10,000 g) for 3 minutes. The supernatant is analyzed byHPLC/MS/MS.

Cytochrome P₄₅₀ Standard CYP1A2 Phenacetin CYP2A6 Coumarin CYP2B6[¹³C]-(S)-mephenytoin CYP2C8 Paclitaxel CYP2C9 Diclofenac CYP2C19[¹³C]-(S)-mephenytoin CYP2D6 (+/−)-Bufuralol CYP2E1 Chlorzoxazone CYP3A4Testosterone CYP4A [¹³C]-Lauric acid

Monoamine Oxidase A Inhibition and Oxidative Turnover

The procedure is carried out using the methods described by Weyler etal., Journal of Biological Chemistry 1985, 260, 13199-13207, which ishereby incorporated by reference in its entirety. Monoamine oxidase Aactivity is measured spectrophotometrically by monitoring the increasein absorbance at 314 nm on oxidation of kynuramine with formation of4-hydroxyquinoline. The measurements are carried out, at 30° C., in 50mM sodium phosphate buffer, pH 7.2, containing 0.2% Triton X-100(monoamine oxidase assay buffer), plus 1 mM kynuramine, and the desiredamount of enzyme in 1 mL total volume.

Monooamine Oxidase B Inhibition and Oxidative Turnover

The procedure is carried out as described in Uebelhack et al.,Pharmacopsychiatry 1998, 31(5), 187-192, which is hereby incorporated byreference in its entirety.

Inhibition of Platelet Aggregation

The procedure is carried out as described in Husted et al., Eur. HeartJ. 2006, 27(9), 1038-1047, which is hereby incorporated by reference inits entirety.

Measuring Pharmacokinetics, and Safety of Ticagrelor

The procedure is carried out as described in Husted et al., EuropeanHeart Journal 2006, 27(9), 1038-1047, which is hereby incorporated byreference in its entirety.

Detecting Ticagrelor and Ticagrelor Metabolites in Humans

The procedure is carried out as described in Butler, et al., Drug MetabRev 2008, 40(Suppl. 3): Abst 280, which is hereby incorporated byreference in its entirety.

Bleeding Time

The procedure is carried out as described in Husted et al., Eur. HeartJ. 2006, 27(9), 1038-1047, which is hereby incorporated by reference inits entirety.

Inhibition of Platelet Aggregation

The procedure is carried out as described in WO 2000034283, which ishereby incorporated by reference in its entirety.

Inhibition of Platelet Aggregation

The procedure is carried out as described in WO 199905142, which ishereby incorporated by reference in its entirety.

From the foregoing description, one skilled in the art can ascertain theessential characteristics of this invention, and without departing fromthe spirit and scope thereof, can make various changes and modificationsof the invention to adapt it to various usages and conditions.

What is claimed is:
 1. A method of treatment of a P2Y12receptor-mediated disorder comprising the administration, to a patientin need thereof, of a therapeutically effective amount of a compoundhaving the structural formula:

wherein each position represented as D has deuterium enrichment of noless than about 10%.
 2. The compound as recited in claim 1 wherein eachposition represented as D has deuterium enrichment of no less than about50%.
 3. The compound as recited in claim 1 wherein each positionrepresented as D has deuterium enrichment of no less than about 90%. 4.The compound as recited in claim 1 wherein each position represented asD has deuterium enrichment of no less than about 98%.
 5. The method asrecited in claim 1 wherein said disorder is selected from the groupconsisting of arterial thrombosis and coronary artery disease.
 6. Themethod as recited in claim 11 further comprising the administration ofan additional therapeutic agent.
 7. The method as recited in claim 6wherein said additional therapeutic agent is selected from the groupconsisting of alpha adrenergic receptor antagonists, beta adrenergicreceptor antagonists, angiotensin II receptor antagonists,angiotensin-converting enzyme inhibitors, anti-arrhythmics,antithrombotics, antiplatelet agents, calcium channel blockers,fibrates, and HMG-CoA reductase inhibitors.
 8. The method as recited inclaim 7 wherein said alpha adrenergic receptor antagonist is selectedfrom the group consisting of, abanoquil, adimolol, ajmalicine,alfuzosin, amosulalol, arotinolol, atiprosin, benoxathian, buflomedil,bunazosin, carvedilol, CI-926, corynanthine, dapiprazole, DL-017,domesticine, doxazosin, eugenodilol, fenspiride, GYKI-12,743,GYKI-16,084, indoramin, ketanserin, L-765,314, labetalol, mephendioxan,metazosin, monatepil, moxisylyte (thymoxamine), naftopidil, nantenine,neldazosin, nicergoline, niguldipine, pelanserin, phendioxan,phenoxybenzamine, phentolamine, piperoxan, prazosin, quinazosin,ritanserin, RS-97,078, SGB-1,534, silodosin, SL-89.0591, spiperone,talipexole, tamsulosin, terazosin, tibalosin, tiodazosin, tipentosin,tolazoline, trimazosin, upidosin, urapidil, zolertine, 1-PP, adimolol,atipamezole, BRL-44408, buflomedil, cirazoline, efaroxan, esmirtazapine,fluparoxan, GYKI-12,743, GYKI-16,084, idazoxan, mianserin, mirtazapine,MK-912, NAN-190, olanzapine, phentolamine, phenoxybenzamine, piperoxan,piribedil, rauwolscine, rotigotine, SB-269,970, setiptiline,spiroxatrine, sunepitron, tolazoline, and yohimbine.
 9. The method asrecited in claim 7 wherein said beta adrenergic receptor antagonist isselected from the group consisting of, acebutolol, adaprolol, adimolol,afurolol, alprenolol, alprenoxime, amosulalol, ancarolol, arnolol,arotinolol, atenolol, befunolol, betaxolol, bevantolol, bisoprolol,bopindolol, bormetolol, bornaprolol, brefonalol, bucindolol, bucumolol,bufetolol, buftiralol, bufuralol, bunitrolol, bunolol, bupranolol,burocrolol, butaxamine, butidrine, butofilolol, capsinolol, carazolol,carpindolol, carteolol, carvedilol, celiprolol, cetamolol, cicloprolol,cinamolol, cloranolol, cyanopindolol, dalbraminol, dexpropranolol,diacetolol, dichloroisoprenaline, dihydroalprenolol, dilevalol,diprafenone, draquinolol, dropranolol, ecastolol, epanolol, ericolol,ersentilide, esatenolol, esmolol, esprolol, eugenodilol, exaprolol,falintolol, flestolol, flusoxolol, hydroxycarteolol, hydroxytertatolol,ICI-118,551, idropranolol, indenolol, indopanolol, iodocyanopindolol,iprocrolol, isoxaprolol, isamoltane, labetalol, landiolol,levobetaxolol, levobunolol, levocicloprolol, levomoprolol, medroxalol,mepindolol, metalol, metipranolol, metoprolol, moprolol, nadolol,nadoxolol, nafetolol, nebivolol, neraminol, nifenalol, nipradilol,oberadilol, oxprenolol, pacrinolol, pafenolol, pamatolol, pargolol,parodilol, penbutolol, penirolol, PhQA-33, pindolol, pirepolol,practolol, primidolol, procinolol, pronethalol, propafenone,propranolol, ridazolol, ronactolol, soquinolol, sotalol, spirendolol, SR59230A, sulfinalol, TA-2005, talinolol, tazolol, teoprolol, tertatolol,terthianolol, tienoxolol, tilisolol, timolol, tiprenolol, tolamolol,toliprolol, tribendilol, trigevolol, xibenolol, and xipranolol.
 10. Themethod as recited in claim 7 wherein said angiotensin II receptorantagonist is selected from the group consisting of candesartan,eprosartan, irbesartan, losartan, olmesartan, tasosartan, telmisartan,and valsartan.
 11. The method as recited in claim 7 wherein saidangiotensin-converting enzyme inhibitor is selected from the groupconsisting of captopril, enalapril, lisinopril, perindopril, ramipril,quinapril, benazepril, cilazapril, fosinopril, trandolapril, spirapril,delapril, moexipril, temocapril, zofenopril, and imidapril.
 12. Themethod as recited in claim 7 wherein said anti-arrhythmic is selectedfrom the group consisting of quinidine, procainamide, disopyramide,sparteine, ajmaline, prajmaline, lorajmine, lidocaine, mexiletine,tocamide, aprindine, propafenone, flecamide, lorcamide, encamide,amiodarone, bretylium tosilate, bunaftine, dofetilide, ibutilidem,tedisamil, moracizine, and cibenzoline.
 13. The method as recited inclaim 7 wherein said antithrombotic is selected from the groupconsisting of dicoumarol, phenindione, warfarin, phenprocoumon,acenocoumarol, ethyl biscoumacetate, clorindione, diphenadione,tioclomarol, heparin, antithrombin III, dalteparin, enoxaparin,nadroparin, parnaparin, reviparin, danaparoid, tinzaparin, sulodexide,bemiparin, ditazole, cloricromen, picotamide, clopidogrel, ticlopidine,acetylsalicylic acid, dipyridamole, carbasalate calcium, epoprostenol,indobufen, iloprost, abciximab, aloxiprin, eptifibatide, tirofiban,triflusal, beraprost, treprostinil, prasugrel, streptokinase, alteplase,urokinase, fibrinolysin, brinase, reteplase, saruplase, ancrod,drotrecogin alfa (activated), tenecteplase, protein C, desirudin,lepirudin, argatroban, melagatran, ximelagatran, bivalirudin, dabigatranetexilate, defibrotide, dermatan sulfate, fondaparinux, and rivaroxaban.14. The method as recited in claim 7 wherein said antiplatelet agent isselected from the group consisting of abciximab, eptifibatide,tirofiban, clopidogrel, prasugrel, ticlopidine, ticagrelor, beraprost,prostacyclin, iloprost, treprostinil, acetylsalicylic acid, aloxiprin,carbasalate calcium, indobufen, dipyridamole, picotamide, terutroban,cilostazol, dipyridamole, triflusal, cloricromen, and ditazole.
 15. Themethod as recited in claim 7 wherein said calcium channel blocker isselected from the group consisting of amlodipine, felodipine,isradipine, nicardipine, nifedipine, nimodipine, nisoldipine,nitrendipine, lacidipine, nilvadipine, manidipine, barnidipine,lercanidipine, cilnidipine, benidipine, mibefradil, verapamil,gallopamil, diltiazem, fendiline, bepridil, lidoflazine, andperhexyline.
 16. The method as recited in claim 7 wherein said fibrateis selected from the group consisting of clofibrate, bezafibrate,aluminium clofibrate, gemfibrozil, fenofibrate, simfibrate, ronifibrate,ciprofibrate, etofibrate, and clofibride.
 17. The method as recited inclaim 7 wherein said HMG-CoA reductase inhibitor is selected from thegroup consisting of atorvastatin, cerivastatin, fluvastatin, lovastatin,mevastatin, pitavastatin, pravastatin, rosuvastatin, and simvastatin.18. The method as recited in claim 1, further resulting in at least oneeffect selected from the group consisting of: a. decreasedinter-individual variation in plasma levels of said compound or ametabolite thereof as compared to the non-isotopically enrichedcompound; b. increased average plasma levels of said compound per dosageunit thereof as compared to the non-isotopically enriched compound; c.decreased average plasma levels of at least one metabolite of saidcompound per dosage unit thereof as compared to the non-isotopicallyenriched compound; d. increased average plasma levels of at least onemetabolite of said compound per dosage unit thereof as compared to thenon-isotopically enriched compound; and e. an improved clinical effectduring the treatment in said subject per dosage unit thereof as comparedto the non-isotopically enriched compound.
 19. The method as recited inclaim 1, further resulting in at least two effects selected from thegroup consisting of: a. decreased inter-individual variation in plasmalevels of said compound or a metabolite thereof as compared to thenon-isotopically enriched compound; b. increased average plasma levelsof said compound per dosage unit thereof as compared to thenon-isotopically enriched compound; c. decreased average plasma levelsof at least one metabolite of said compound per dosage unit thereof ascompared to the non-isotopically enriched compound; d. increased averageplasma levels of at least one metabolite of said compound per dosageunit thereof as compared to the non-isotopically enriched compound; ande. an improved clinical effect during the treatment in said subject perdosage unit thereof as compared to the non-isotopically enrichedcompound.
 20. The method as recited in claim 1, wherein the methodeffects a decreased metabolism of the compound per dosage unit thereofby at least one polymorphically-expressed cytochrome P₄₅₀ isoform in thesubject, as compared to the corresponding non-isotopically enrichedcompound.
 21. The method as recited in claim 20, wherein the cytochromeP₄₅₀ isoform is selected from the group consisting of CYP2C8, CYP2C9,CYP2C19, and CYP2D6.
 22. The method as recited claim 1, wherein saidcompound is characterized by decreased inhibition of at least onecytochrome P₄₅₀ or monoamine oxidase isoform in said subject per dosageunit thereof as compared to the non-isotopically enriched compound. 23.The method as recited in claim 22, wherein said cytochrome P₄₅₀ ormonoamine oxidase isoform is selected from the group consisting ofCYP1A1, CYP1A2, CYP1B1, CYP2A6, CYP2A13, CYP2B6, CYP2C8, CYP2C9,CYP2C18, CYP2C19, CYP2D6, CYP2E1, CYP2G1, CYP2J2, CYP2R1, CYP2S1,CYP3A4, CYP3A5, CYP3A5P1, CYP3A5P2, CYP3A7, CYP4A11, CYP4B1, CYP4F2,CYP4F3, CYP4F8, CYP4F11, CYP4F12, CYP4X1, CYP4Z1, CYP5A1, CYP7A1,CYP7B1, CYP8A1, CYP8B1, CYP11A1, CYP11B1, CYP11B2, CYP17, CYP19, CYP21,CYP24, CYP26A1, CYP26B1, CYP27A1, CYP27B1, CYP39, CYP46, CYP51, MAO_(A),and MAO_(B).
 24. The method as recited in claim 1, wherein the methodreduces a deleterious change in a diagnostic hepatobiliary functionendpoint, as compared to the corresponding non-isotopically enrichedcompound.
 25. The method as recited in claim 24, wherein the diagnostichepatobiliary function endpoint is selected from the group consisting ofalanine aminotransferase (“ALT”), serum glutamic-pyruvic transaminase(“SGPT”), aspartate aminotransferase (“AST,” “SGOT”), ALT/AST ratios,serum aldolase, alkaline phosphatase (“ALP”), ammonia levels, bilirubin,gamma-glutamyl transpeptidase (“GGTP,” “γ-GTP,” “GGT”), leucineaminopeptidase (“LAP”), liver biopsy, liver ultrasonography, livernuclear scan, 5′-nucleotidase, and blood protein.